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LAPORAN PRAKTIKUM : ISOLASI DNA MAY 29, 2014 / ASTRIDSAFIRAIDHAM
LAPORAN PRAKTIKUM GENETIKA PERCOBAAN II ISOLASI DNA NAMA
: ASTRID SAFIRA IDHAM
NIM
: H41113341
HARI/TANGGAL KELOMPOK ASISTEN
: KAMIS, 13 MARET 2014 : VI (ENAM) B : ANDRI NIND NINDA A
LABORATORIUM GENETIKA !URUSAN BIOLOGI FAKULTAS MATEMATIKA DAN ILMU PENGETAHUAN ALAM UNIVERSITAS HASANUDDIN MAKASSAR 2014
BAB I PENDAHULUAN I"1 L#$#% B&'##* Deoxyribonucleic Deoxyribonucle ic acid atau DNA meu!a"a# $e#%a&a "'m'a %a#(
!a)'#( !e#t'#( *a)am ma"+)u" +'*u!DNA meu!a"a# $e#%a&a %a#( me#(a#*u#( '#-.ma$' (e#et'" ma"+)u" +'*u! *a' $atu (e#ea$' "e (e#ea$' $e)a#ut#%a Ke$e)uu+a# DNA *a)am $uatu $e) a"a# meme#tu" (e#.m e#.m me)'!ut' a('a# (e# %a#( -u#($'.#a) mau!u# #.#-u#($'.#a) *a)am $e) .(a#'$me DNA (e#.m me)'!ut' (e# *a#'#te(e# 3Su%., 2004 DNA a*a)a+ a$am #u")eat %a#( me#(a#*u#( mate' (e#et'" *a# e-u#($' u#tu" me#(atu !e"ema#(a# '.).('$ $e)uu+ e#tu" "e+'*u!a# $e5aa $e)u)e DNA te*a!at !a*a #u")eu$, m't'".#*'a, *a# ")..!)a$ Pee*aa# "et'(a#%a a*a)a+ DNA #u")eu$ ee#tu" )'#'e *a# ea$.$'a$' $a#(at eat *e#(a# !.te'# +'$t.#, $e*a#("a# DNA m't.".#*'a *a# ")..!)a$ ee#tu" $'"u)a *a# t'*a" ea$.$'a$' *e#(a# !.te'# +'$t.# Se)a'# 'tu DNA m't.".#*'a *a# ")..!)a$ mem')'"' mem')'"' 5'' "+a$, %a'tu +a#%a me&a'$"a# $'-at$'-at %a#( ea$a) *a' (a'$ 'u Se*a#("a# DNA #u")eu$ mem')'"' !.)a !e&a'$a# $'-at *a' "e*ua .a#(tua D')'+at *a' .(a#'$me#%a, $tu"tu DNA !."a'.t t'*a" mem')'"' !.te'# +'$t.# *a# ee#tu" $'"u)a, $e*a#("a# DNA eu"a'.t ee#tu" )'#'e *a# mem')'"' !.te'# +'$t.# 3K'$ma#, 2010 I$.)a$' DNA meu!a"a# )a#("a+ mem!e)aa' DNA Sa)a+ $atu !'#$'!'$.)a$' !'#$'!'$ .)a$' DNA %a'tu *e#(a# $e#t'-u(a$' Se#t'-u(a$' meu!a"a# te"#'" u#tu"mem'$a+"a# 5am!ua# e*a$a"a# eat
m.)e"u) ".m!.#e##%a M.)e"u) %a#( mem!u#%a' eat m.)e"u) e$a a"a# ea*a *' a('a# a&a+ tau#( *a# m.)e"u) '#(a# a"a# ea*a !a*a a('a# ata$ tau#( Ha$') $e#t'-u(a$' a"a# me#u#u""a# *ua ma5am -a"$' %a#( te!'$a+, %a'tu $u!e#ata# !a*a a('a# ata$ *a# !e)et !a*a a('a# a&a+ 3Ra5+mat, 2012
I"2 T++# P&%-.##
A*a!u# tuua# %a#( a"a# *'5a!a' !a*a !e5.aa# '$.)a$' DNA '#' %a'tu u#tu" me#(eta+u' 5aa mem'$a+"a# 3me#('$.)a$' DNA *a' 2 e#'$ ua+ %a#( *'a*'"a# $ea(a' $am!e), $eta u#tu" me#(eta+u' "ea"t'-a# *ete(e# *a)am !.$e$ '$.)a$' DNA !a*a $am!e) ua+
I"3 #$+ # T&#$ P&%-.##
Pe5.aa# '#' *')a"$a#a"a# !a*a +a' Kam'$, 16 Maet 2014, !u"u) 1400 7 1860 ITA Pe5.aa# '#' etem!at *' :a.at.'um B'.).(' Da$a, ;uu$a# B'.).(', Fa"u)ta$ Matemat'"a *a# I)mu Pe#(eta+ua# A)am, U#'
BAB II TIN!AUAN PUSTAKA
I$.)a$' DNA a*a)a+ !.$e$ !e#(e)uaa# DNA *a' tem!at#%a ea*a 3e"$ta"$' atau )'$'$ 'a$a#%a *')a"u"a# *e#(a# +.m.(e#a$' *a# !e#ama+a# u=e e"$ta"$' atau u=e )'$'$ u#tu" me#5e(a+ DNA u$a" 3Yu&.#., 200> Pa*a $e) eu"a'.t'" tema$u" ta#ama# *a# +e&a# a('a# tee$a *a' DNA ea*a !a*a #u")eu$ %a'tu .(a#e) %a#( *'!'$a+"a# *a' $'t.!)a$ma *e#(a# mema# Nu")eu$ te*'' *a' 90 ? "e$e)uu+a# DNA $e)u)e S'$a DNA a*a)a+ .(a#e) )a'# $e!et' m't.".#*'a *a# ")..!)a$ Kae#a DNA te*a!at !a*a #u")eu$, ma"a !e)u a*a#%a met.*e !e)'$'$a# $e) $am!a' !ema#e#a# $e) D'ma#a met.*e te$eut meu!a"a# a('a# *a' met.*e '$.)a$' DNA 3E).*, 2008 Pem'$a+a# DNA *a' mate' $e)u)e )a'##%a meu!a"a# +a) %a#( $'(#'@"a# *a# me#(+au$"a# !e#%a))'#a# DNA me#a*' RNA *a# ta#$)a$' RNA me#a*' !.te'# e)a#($u#( *a)am ".m!ateme#3 ua#( %a#( ee*a %a'tu $e5aa etuuttuut *a)am #u5)eu$ *a# $'t.!)a$ma 3 E).*, 2008
Te*a!at .(a#e).(a#e) emema# (a#*a *' *a)am $'t.!)a$ma, tema$u" m't.".#*'a a'" !a*a tumu+a# mau!u# +e&a# O)e+ "ae#a 'tu !e)u *')a"u"a# '$.)a$' DNA !a*a ta#ama# *a# +e&a# u#tu" me#(eta+u' *a# mem!e)aa' DNA *a' ta#ama# *a# +e&a# te$eut Se) eu"a'.t'" mem')'"' DNA )e'+ a#%a", )e#("a! *e#(a# ".m!.#e#".m!.#e# )a'# DNA ta#ama# *a# +e&a# te$'m!a# *a)am #u5)eu$%a#( teu#("u$ .)e+ mema# I$.)a$' DNA meu!a"a# )a#("a+ %a#( te!at u#tu" mem!e)aa' DNA P'#$'!#%a a*a *ua, %a'tu $e#t'-u(a$' *a# !e$'!'ta$'Se#t'-u(a$' meu!a"a# te"#'" u#tu" mem'$a+"a# 5am!ua# e*a$a"a# eat m.)e"u) ".m!.#e##%a M.)e"u) %a#( mem!u#%a' eat m.)e"u) e$a a"a# ea*a *' a('a# a&a+ tau#( *a# m.)e"u) '#(a# a"a# ea*a !a*a a('a# ata$ tau#( Ha$') $e#t'-u(a$' a"a# me#u#u""a# *ua ma5am -a"$' %a#( te!'$a+, %a'tu $u!e#ata# !a*a a('a# ata$ *a# !e)et !a*a a('a# a&a+ Pe$'!'ta$' meu!a"a# )a#("a+ %a#( *')a"u"a# u#tu" me#(e#*a!"a# $uatu ".m!.#e# *a' 5am!ua# 3A)et, 1994 DNA !a*a ma"+)u" +'*u! *a!at *''$.)a$' $e5aa $e*e+a#a Pe#('$.)a$'a# DNA $e5aa $e*e+a#a *a!at *')a"u"a# *e#(a# meme5a+"a# *'#*'#( $e), mema# !)a$ma *a# mema# '#t' a'" $e5aa me"a#'" mau!u# $e5aa "'m'a&' I$.)a$' DNA meu!a"a# $uatu te"#'" %a#( *'(u#a"a# u#tu" mem!e.)e+ DNA mu#', %a'tu ta#!a !.te'# *a# RNA *a' $uatu $e) *a)am a'#(a# Peme5a+a# *'#*'#( $e) $e5aa me"a#'" *a!at *')a"u"a# *e#(a# !em)e#*ea# atau !e#((eu$ me#((u#a"a# m.ta *a# !'$t') Se*a#("a# $e5aa "'m'a&' *a!at *')a"u"a# *e#(a# !eme'a# *ete(e# Pe#ama+a#
$au# 5a' *a# (aam *a!u a*a)a+ u#tu" me)'$'$"a# mema# '#t' u#tu" me#(e)ua"a# '$' '#t' $e) %a#( e'$' DNA 3Ra5+mat, 2012 I$.)a$' DNA *a!at *')a"u"a# me)a)u' ta+a!a#ta+a!a# a#taa )a'# !e!aa$' e"$ta" $e), !emu#'a# DNA *a' e"$t$" $e) *a# !e$'!'ta$' DNA Me$"'!u# '$.)a$' DNA *a!at *')a"u"a# *e#(a# ea(a' 5aa, a"a# teta!' !a*a $et'a! e#'$ atau a('a# ta#ama# *a!at meme'"a# +a$') %a#( ee*a, +a) '#' *'"ae#a"a# a*a#%a $e#%a&a !.)'-e#.) *a# !.)'$a"a'*a *a)am ".#$e#ta$' t'#((' %a#( *a!at me#(+amat !emu#'a# DNA ;'"a '$.)a$' DNA *')a"u"a# *e#(a# $am!)e ua+, ma"a "a*a a' !a*a ma$'#(ma$'#( ua+ ee*a, *a!at meme' +a$') %a#( ee*ae*a !u)a Sema"'# t'#((' "a*a a', ma"a $e) %a#( te)aut *' *a)am e"$ta" a"a# $ema"'# $e*'"'t, $e+'#((a DNA %a#( te!et'$'!a$' u(a a"a# $e*'"'t 3K'$ma#, 2010 Pe#ama+a# *etee# *a)am '$.)a$' DNA *a!at me#%ea"a# u$a"#%a mema#e $e), me)a)u' '"ata# %a#( *'e#tu" me)a)u' $'$' +'*.-.'" *etee# *e#(a# !.te'# *a# )ema" !a*a mema#e meme#tu" $e#%a&a )'!'* !.te'#*etee# ".m!)e"$ Se#%a&a te$eut *a!at tee#tu" "ae#a !.te'# *a# )'!'* mem')'"' uu#( +'*.@)'" *a# +'*.-.'", *em'"'a# u(a *e#(a# *etee#, $e+'#((a *a!at meme#tu" $uatu '"ata# "'m'a 3K'$ma#, 2010 P'#$'! 7 !'#$'! *a)am me#('$.)a$' DNA 3T.+', 2012 1 me)'$'$ $e) $e5aa @$'", *e#(a# 5aa !e#((eu$a# 2 !eme5a+a# *'#*'#( $e)
6 !eme5a+a# mema# $e) 4 !em'$a+a# DNA *a' a+a# %a#( )a'# Da)am '$.)a$' DNA, a+a# %a#( "'ta (u#a"a# 'a$a#%a eu!a a'#(a# tumu+a# atau a'#(a# +e&a#, u#tu" 'tu )a#("a+ !etama %a#( +au$ "'ta )a"u"a# a*a)a+ meme5a+"a# a'#(a# me#a*' $e)$e) %a#( ma#*'' P.$e$ *')a"u"a# $e5aa me"a#'" atau @$'" *e#(a# me#umu" atau me#((eu$ a+a# %a#( a"a# "'ta (u#a"a# *e#(a# m.ta atau )e#*e Ke*ua a*a)a+ meme5a+"a# *'#*'#( $e) *a# mema# $e) )a!'$a# !emu#("u$ DNA $tu"tu utama !eme#tu" mema# *a# *'#*'#( $e) a*a)a+ )ema", u#tu" 'tu "'ta (u#a"a# *etee# *a# (aam *a!u Ke*ua a+a# '#' *'(u#a"a# u#tu" me)ua#(' *a# meu$a" $e) $e+'#((a '$' '#t' $e) 3DNA '$a "e)ua 3T.+', 2012 Ta+a! $e)a#ut#%a a*a)a+ !em'$a+a# DNA *a' a+a# %a#( )a'# Pem'$a+a# *')a"u"a# *e#(a# me#((u#a"a# et+a#.)/a)".+.) *'#('# e".#$e#ta$' 909? Et+a#.)/A)".+.) t'*a" me)aut"a# DNA *a# eat e#'$ a)".+.) %a#( )e'+ '#(a# *a' a' memuat DNA #a'" *a# me)a%a#()a%a#( *' !emu"aa# 3T.+', 2012 CTAB atau Cetyl trimethylammonium bromide meu!a"a# $ee#'$ *etee# %a#( *a!at me#*e(a*a$' *'#*'#( $e), *e#atua$' !.te'#, mem'$a+"a# "a.+'*at, meu$a" mema# $e) *a# me)aut"a# DNA A!a')a *'#*'#( $e) te*e(a*a$' ma"a $emua '$' $e) *a!at "e)ua tema$u" DNA *a# *')e!a$"a# "e *a)am u=e e"$ta"$'Da)am !.$e$ '$.)a$' DNA ta#ama#, !e#ama+a# $e#%a&a !ee*u"$' $e!et' me5+a!t.eta#.) *a!at me#5e(a+
!.$e$ ."$'*a$' $e#%a&a -e#.)'" $e+'#((a me#(+amat a"t'<'ta$ a*'"a) ea$ %a#( *'+a$')"a# .)e+ ."$'*a$' -e#.) te+a*a! a$am #u")eat Me5+a!t.eta#.) u(a e-u#($' u#tu" me)'#*u#(' RNA *a' $e#%a&a u'#.#, *'$u)!+'*e, !e."$'*a, !.)'!+e#."$'*a$e, *a# !.te'# P.$e$ !ema#$a# !etama etuua# u#tu" me)aut"a# CTAB *a# me5a!t.eta#.) Se*a#("a# !ema#a$a# %a#( "e*ua etuua# u#tu" mem*e(a*a$' !.te'# *a# *'#*'#( $e) 3T.+', 2012 Klorofrom dan isoamilalkohol 3CIAA e-u#($' u#tu" me#(e"$ta"
*a# *a# me#(e#*a!"a# ".m!.#e# !.)'$a"a'*a *' *a)am u=e e"ta"$' %a#( me#(".#tam'#a$' )auta# DNA Peme'a# '$.!.!a#.) *a# eta#.) *')a"u"a# a(a tea*' *e+'*a$' DNA $e+'#((a tea*' !e$'!'ta$' Sete)a+ !eme'a# eta#.), !e))et %a#( *'!e.)e+ *'"e'#(a#('#"a# Ha) '#' etuua# u#tu" me#(e'#("a# !e))et *a' $'$a$'$a u=e mau!u# eta#.)Ta+a!a# tea"+' *a' e"ta"$' '#' a*a)a+ !e#ama+a# u=e TE Bu=e TE tris electrophoresise-u#($' u#tu" me)aut"a# DNA %a#(*'+a$')"a# *a#
me#a(aDNA a(a t'*a" mu*a+ u$a" Da)am u=e TE me#(a#*u#( EDTA atau Ethylene Diamine Tetra Acid %a#( e-u#($' $ea(a' $e#%a&a !e#("e)at %a#( me#('"at '.# Ma(#e$'um, %a'tu ".-a"t. %a#( *'!e)u"a# u#tu" a)t'
CTAB u(a a"a# *'!e.)e+ RNA *e#(a# !'ta t'!'$ %a#( te)eta" au+ ea*a *' a&a+ !'ta DNA Keea*aa# !'ta RNA te(a#tu#( a+a# %a#( *'e"$ta"$' 3A$'$, 2010
Met.*e '#' t'*a" memutu+"a# 'a%a %a#( )e'+ ma+a) *'a#*'#("a# *e#(a# me#((u#a"a# "'t Se)a'# 'tu, "e)e'+a# *a' e"ta"$' '#' a*a)a+ !'ta DNA %a#(*'!.)e+ )e'+ tea) ')a *'a#*'#()a# *e#(a# e"ta"$' met.*e -e#.) *a# ta#!a -e#.) A"a# teta!', *a' +a$') *e#(a# met.*e '#' ma$'+ te*a!at !'ta $mea *a# DNA %a#( *'+a$')"a# )e'+ $e*'"'t *a'!a*a e"ta"$' *e#(a# me#((u#a"a# "'t Ke#*a)a %a#( umum tea*' *a)am e"$ta"$' CTAB a*a)a+ a*a#%a '#+''t. !a*a '#a#(, e#*a+#%a ".#$e#ta$' <'u$ *a# !e#(au+ 5aa mau!u# )ama &a"tu !e#%'m!a#a# 3A$'$, 2010
BAB III
METODE PERCOBAAN III"1 A'#$ # B## III"1"1 A'#$
A)at 7 a)at %a#( *'(u#a"a# *a)am !e5.aa# '#' a*a)a+ a)at tu)'$ me#u)'$, tau#( ea"$', t'ma#(a#, (e)a$ aua, !'$au, ata#( !e#(a*u", $!atu)a, )e#*e, !'!et tete$, *a# me$'# <.teG III"1"2 B##
Ba+a# %a#( *'(u#a"a# *a)am !e5.aa# '#' a*a)a+ ua+ t.mat Solanum lycopercium *a# !e!a%a Carica papaya, *ete(e# 3R'#$. 5a', Su- uu", *a# $au# u"'m, Aua*e$, (aam *a!u, Et+a#.) 9 ? , *a# "eta$ $a'#(
III"2 P%.5&+% K&%#
A*a!u# )a#("a+)a#("a+ "ea %a#( *')a"u"a# *a)am !e5.aa# '#' a*a)a+ $ea(a' e'"ut 1 Bua+ *'!.t.#( me#a*' u"ua# %a#( )e'+ "e5') 2 Bua+ %a#( te)a+ *'!.t.#( "emu*'a# *'t'ma#( $am!a' eat#%a 100 ( 6 D'am') 100 ( ua+ *a# 100 m) aua*e$, "emu*'a# *')e#*e +'#((a +a)u$, "'a"'a *')e#*e $e)ama 40 *et'" 4 Ma$'#(ma$'#( ua+ %a#( te)a+ *')e#*e *'ma$u""a# "e*a)am 6 tau#( ea"$' %a#( ee*a $ea#%a" 4 m) D')aut"a# *ete(e# 3R'#$., Su-, *a# Bu"'m "e*a)am 0 m) aua*e$, "emu*'a# *'a*u" +'#((a )aut
T'(a e#'$ )auta# *ete(e# "emu*'a# *'ma$u""a# "e*a)am ma$'#(ma$'#( tau#( ea"$' %a#( e'$' u$ ua+ $ea#%a" 4 m) 8 Tama+"a# 1 $!atu)a (aam *a!u "e*a)am ma$'#(ma$'#( tau#( ea"$', "emu*'a# *'a*u" +'#((a )aut > Cam!ua# "emu*'a# *'$a'#( $ea#%a" 2 "a)' *e#(a# "eta$ $a'#( 9 :a)u *'tama+"a# et+a#.) 9 ? $ea#%a" m) "e*a)am ma$'#(ma$'#( 5am!ua# 10
:auta# "emu*'a# *'+.m.(e#"a# *e#(a# me#((u#a"a#
me$'# <.teG 11
Amat' *a# 5atat +a$') %a#( tam!a" *a' "e$e)uu+a#
!.$e$, me)'!ut' &a#a, $eta $e*'"'t a#%a"#%a DNA %a#( tee#tu"
DAFTAR PUSTAKA
A)et, B, 1994 Biologi olekuler Sel Edisi Kedua PT ame*'a Pu$ta"a Utama ;;;;;;;a"ata
A$'$, 2010 De!nisi "solasi D#A dan anfaat "solasi D#A+tt!//a$'$08Stu*e#t'!a5'* D'a"$e$ !a*a ta#((a) 18 Maet
2014, !a*a !u"u) 2600 ITA E).*, S, 2008 $enetika Edisi Keempat E)a#((a ;a"ata K'$ma#, 2010"solasi D#A Buah +tt!//"'$ma#>6&ee) 5.m D'a"$e$ !a*a ttttttttta#((a) 18 Maet 2014, !a*a !u"u) 2600 ITA
Ra5+mat, 2012 "solasi D#A% +tt!//a5+mat%u$u-).($!.t5.m D'a"$e$ !a*a $$$$$$$ta#((a) 18 Maet 2014, !a*a !u"u) 2600 ITA
Su%., 2004 $enetika Strata &% U#'
Yu&.#., T, 200> Biologi olekuler E)a#((a ;a"ata
BAB IV HASIL DAN PEMBAHASAN IV"1 H#56' P&%-.## IV"1"1 G##% H#56' P&%-.## !ENIS BUAH
SEBELUM
SESUDAH
1"PEPAA Carica papaya
(++, -#6%, %6)
(++, -#6%, %6)
2"T.#$Solanu m lycopercium
(%6, ++, -#6%)
(++, -#6%, %6)
IV"1"2 T#&' P&*##$# HASIL PERCOBAAN !ENIS BUAH
PERLAKUAN
ARNA
AKTU
!UMLAH
Dete(e# Buu"
Put'+ 7 e#'#(
Ce!at
Dete(e# Ca'
Ku#'#( 7 e#'#(
:amat
Dete(e# K'm
Ku#'#( 7 !u5at
Se*a#(
Dete(e# Buu"
Put'+ 7 e#'#(
Ce!at
Dete(e# Ca'
Be#'#(
:amat
Dete(e# K'm
Ku#'#(
Se*a#(
Pe!a%a Carica papaya
T.mat Solanum lycoperciu m
IV"2 P&##5#
P'#$'! utama *a)am '$.)a$' DNA a*a t'(a %a"#' !e#(+a#5ua# 3)'$'$, e"ta"$' atau !em'$a+a# DNA *a' a+a# !a*at $e!et' $e)u).$a *a# !.te'#, $eta !emu#'a# DNA Se)a'# 'tu a*a 2 !'#$'! )a(' *a)am me)a"u"a# '$.)a$' DNA, %a'tu $e#t'-u(a$' *a# !e$'!'ta$' P'#$'! utama $e#t'-u(a$' a*a)a+ mem'$a+"a# $u$ta#$' e*a$a"a# eat e#'$ m.)e"u) *e#(a# 5aa meme'"a# (a%a $e#t'-u(a) $e+'#((a $u$ta#$' %a#( )e'+ eat a"a# ea*a *' *a$a, $e*a#("a# $u$ta#$' %a#( )e'+ '#(a# a"a# te)eta" *' ata$ Te"#'" $e#t'-u(a$' te$eut *')a"u"a# *' *a)am $eua+ me$'# %a#( e#ama me$'# $e#t'-u(a$' *e#(a# "e5e!ata# %a#( e
tee#tu" a#taa *ete(e# *e#(a# atat %a#( a*a !a*a ua+ Sete)a+ !e#ama+a# *ete(e# ta+a! $e)a#ut#%a %a'tu !e#ama+a#a (aam *a!u $ea#%a" 1 $!atu)a "e ma$'#(ma$'#( tau#( %a#( e'$' 5am!ua# ua+ *a# *ete(e#, tuua# *a' !e#ama+a# (aam a*a)a+ u#tu" memu*a+"a# !em'$a+a# e#a#( e#a#( DNA *a' 5am!ua# $e+'#((a e#a#(e#a#( te$eut a"a# mu*a+ *'amat' Ha) '#' tea*' "ae#a Na *a)am (aam *a!at meme#tu" '"ata# !a*a "utu #e(at'
mau!u# "a#*u#(a# a' *a' ma$'#(ma$'#( ua+ t'*a" $ama Da' !e5.aa# t'*a" *'!e.)e+ DNA mu#' me)a'#"a# +a#%a DNA "a$a atau e#a#( 7 e#a#( +a)u$ 3$u!e#ata#t *a)am um)a+ %a#( $e*'"'t $eta e#*a!a# !ut'+ %a#( te)'+at Da' "e*ua ua+ DNA "a$a %a#( !a)'#( te)'+at a*a !a*a ua+ !e!a%a, +a) 'tu tea*' a"'at "a#*u#(a# a' !e!a%a )e'+ $e*'"'t *a' t.mat $e+'#((a e"$ta" *a# DNA "a$a %a#( *'+a$')"a# .)e+ !e!a%a )e'+ a#%a" *a'!a*a ua+ t.mat, $e)a'# 'tu e#*a!a# !ut'+ !a*a ua+ !e!a%a )e'+ a#%a" *a'!a*a ua+ t.mat
BAB V PENUTUP V"1 K&56+'#
Da' !e5.aa# '$.)a$' DNA %a#( te)a+ *')a"u"a# *a!at *'"eta+u' met.*e atau 5aa a(a'ma#a me#('$.)a$' DNA *a' ua+ %a#( *'a*'"a# $ea(a' $am!e) Ya'tu me)a)u 5aa !e#(+a#5ua# 3)'$'$, e"$ta"$', $eta !emu#'a# Pe#((u#aa# *ete(e# %a#( ee*a *a)am !e5.aa# *')a"u"a# u#tu" me#(eta+u' *ete(e# *a)am e#tu" a!a %a#( !a)'#( a"t'- *a)am me#(+a#5u"a# mema# $e) !a*a $et'a! $am!e) ua+Sete)a+ *')a"u"a# !e5.aa# te#%ata *ete(e# uu" me#(+a$')"a# DNA "a$a %a#( )e'+ te)'+at $eta e#*a!a# !ut'+ %a#( )e'+ a#%a"
V"2 S#%# V"2"1 S#%# U$+ L#.%#$.%6+
Sea'"#%a )a.at.'um me#%e*'a"a# a)at %a#( 5u"u! mema*a' $eta a)at %a#( 5u"u! a#%a", a(a !a"t'"um *a)am ea)a# *e#(a# e@$'e# V"2"2 S#%# U$+ A565$&
D'+aa!"a# a(a teu$ mem!e+at'"a# *a# mem'm'#( !a"t'"a# $e)ama )a e)a#($u#(, a(a !a"t'"um ea)a# *e#(a# a'"
Ade puji setyawati Biologi inspirasi Jumat, 14 November 2014
!"#A! $NA %ANA&AN ISOLASI DNA TANAMAN
A*e Pu' Set%a&at' 1 D+ B+'#tat' S Ha$ta', M B'.me* 2 Fe$t% Au)'%au Ra+ma+, S S' 2 Nu(.+. A*' Mau)a#a 6 I#*+'#a Re'+a##'$+a 6 1
Ma+a$'$&a ;uu$a# B'.).(', Fa"u)ta$ Sa'#$ *a# Te"#.).(', U#'
D.$e# Pa"t'"um B'.).(' M.)e"u)e
6
A$'$te# Pa"t'"um B'.).(' M.)e"u)e
;) I H Dua#*a, Ta#(ea#( Se)ata# 1412, I#*.#e$'a Ema') A*e'.12J(ma')5.m ;umat, 24 O"t.e 2014
ABSTRAK D#A merupakan senya'a yang mengandung informasi genetik makhluk hidup dari satu generasi ke generasi selan(utnya% D#A pada sel tanaman dibagi men(adi dua yaitu D#A intrakromosomal dan D#A ekstrakromosomal% )risnsip utama dalam isolasi D#A ada tiga yakni penghancuran *lisis+, ektraksi atau pemisahan D#A dari dinding sel, membrane sel dan protein, serta pemurnian D#A% "solasi D#A sa'i hi(au ini dilakukan dengan mengambil potongan daun muda kemudian daun tersebut diekstraksi hingga didapatkan D#A murni dari tanaman
sa'i hi(au tersebut% )raktikum ini dilakukan dengan tu(uan agar praktikan mampu melakukan isolasi D#A dari tanaman Sa'i -i(au *Brassica sp%+% -asil setelah penambahan kloroform dan isoamil alkohol *./0&+ &x 1olum sampel dan disentrifugasi terbentuk . fase yaitu fase a2oueous dan organik% Setelah penambahan isopropanol dingin terlihat benang3benang tipis D#A%
Kata kunci 0 Tanaman, daun Sa'i hi(au, pemurnian D#A
I"
D#5#% T&.%6
De.G%'. Nu5)e'5 a5'* 3DNA meu!a"a# $e#%a&a "'m'a %a#( !a)'#( !e#t'#( *a)am ma"+)u" +'*u! DNA meu!a"a# $e#%a&a %a#( me#(a#*u#( '#-.ma$' (e#et'" ma"+)u" +'*u! *a' $atu (e#ea$' "e (e#ea$' $e)a#ut#%a DNA eu"a'.t t'*a" +a#%a *'um!a' !a*a #u")eu$, teta!' *a!at *'temu"a# !a*a m't.".#*'a *a# ")..!)a$ DNA %a#( *''$.)a$' *a' ")..!)a$ me#u#u""a# $'-at ee#tu" $'"u)a, te*'' *a' u#ta' (a#*a, e!)'"a$' $em'".#$e Met.*e te$eut !a*a *a$a#%a mem')'"' !'#$'! %a#( $ama, #amu# a*a eea!a +a) tete#tu %a#( 'a$a#%a *'(u#a"a# m.*'@"a$' u#tu" *a!at me#(+a#5u"a# '#+''t. %a#( a*a *' *a)am ma$'#(ma$'#( $ume $!e$'me# K.m!.#e# #u")e.t'*a DNA a*a)a+ (u)a, -.$-at, *a# a$a #'t.(e# K.m!.#e# (u)a !a*a DNA a*a)a+ (u)a *e."$''.$a, %a'tu (u)a '.$e %a#( "e+')a#(a# $atu at.m ."$'(e# Ba$a %a#( a*a !a*a DNA a*a *ua ma5am, %a'tu !u'# *a# !''m'*'# Pu'# tea(' )a(' me#a*' *ua ma5am, %a'tu a*e#'# *a# (ua#'# P''m'*'# te*'' *a' *ua e#'$, %a'tu t'm'# *a#$'t.$'# 3 Yu&.#., 200> Nu")eu$ te*'' *a' 90 ? "e$e)uu+a# DNA $e)u)e S'$a DNA a*a)a+ .(a#e) )a'# $e! et' m't.".#*'a *a# ")..!)a$ 3He)*t, 200 Kae#a DNA te*a!at !a*a #u")eu$, ma"a !e)u a*a#%a met.*e !e)'$'$a# $e) $am!a' !ema#e#a# $e) Sa&' +'au 3 Brassica sp% meu!a"a# e#'$ $a%u %a#( *'(ema' .)e+ ma$%aa"at I#*.#e$'a Ke)e'+a#"e)e'+a# $a&' a#taa )a'# a'" a(' "e$e+ata# tuu+, mam!u tumu+ a'" a'" *' *ataa# e#*a+ mau!u# *ataa# t'#((', ta+a# te+a*a! a' +ua#, *a!at *'!a#e# $e!a#a#( ta+u# t'*a" te(a#tu#( *e#(a# mu$'m, ma$a !a#e##%a 5u"u! !e#*e", %a'tu $e"'ta 40 +a' $ete)a+ ta#am *a# $a&' mem!u#%a' #')a' e".#.m' 5u"u! t'#((' $ete)a+ "u'$ ".!, "u'$ u#(a, *a# .".)' 3A*'a#a, 2009
De#(a# me#(eta+u' DNA ta#ama# $a&' ma"a *a!at me#(+a$')"a# !.te#$' %a#( 5u"u! e$a u#tu" me#(+a$')"a#
A)at %a#( *'(u#a"a# *a)am !a"t'"um '$.)a$' DNA ta#ama# a*a)a+ a)at tu)'$, m'".!'!et, $e#t'-u(a, <.teG, &ateat+, '#"uat., yello' tips, blue tips, -eee,t'! $te'), tau#( m'". 1, $te'), (u#t'#(, t'ma#(a# a#a)'t'" *a# "amea Ba+a# %a#( *'(u#a"a# a*a)a+ *au# $a&' +'au *Brassica sp%+% %a#( ma$'+ mu*a 0,6 (, 00 L: u=e e"$ta"$' 32 mM EDTA, 20 mM NaC), SDS 0,?, 200 mM T'$HC) !H 8,, ")..-.m *a# '$.am') a)".+.) 3241 $ea#%a" 1G <.)um $am!e), '$.!.!a#.) $ea#%a" 1G <.)ume $am!e), *a# 100 L: u=e TE III"
M&$.&
Caa "ea u#tu" '$.)a$' DNA ta#ama# %a'tu !etama *e#(a# me#'ma#( *au# $a&' +'au %a#( ma$'+ $e(a $ea#%a" 0,6 (am, ta#!a !etu)a#(a# *au# *a# ata#(#%a Kemu*'a# *'ma$u"a# "e*a)am tau#( m'". $te') Sete)a+ 'tu )u#a"a# *au# $e)ama 10 me#'t !a*a $u+u ua#(, (u#a"a# t'! $te') u#tu" me#umu" *au#, ta#!a !e#ama+a# u--e *a# tama+"a# 00 : u-e e"$ta"$' 3200 mM T'$HC) 3!H 8, 20 mM NaC), 2 mM EDTA, 0? SDS "e *a)am tau#( Kemu*'a# ".5." $e)ama *et'" "emu*'a# I#"ua$' tau#( *a)am &ateat+ !a*a $u+u 00C $e)ama 60 me#'t *e#(a# tamaa+a# ")..-.m'$.am')a)".+.) 3241$ea#%a" 1G <.)ume $am!e) )a)u ".5." "emu*'a# $e#t'-u(a !a*a 000 G ( $e)ama me#'t *a# *' am') $u!e#ata#, "emu*'a# !'#*a+"a# "e tau#( m'". %a#( au, )a)u tama+"a# '$.!.!a#.) $ea#%a" 1G <.)ume $am!e), K.5." "emu*'a# '#"ua$' !a*a $u+u 20 0C $e)ama 1 me#'t Se#t'-u(a$' !a*a 000 G ( $e)ama me#'t, $ete)a+ 'tu $u!e#ata#, "emu*'a# *'tama+"a# 100 : u-e TE *a# $'m!a# !a*a $u+u 20 0C IV"
H#56' # P&##5#
Ha$') %a#( *'*a!at"a# *a' !a"t'"um '$.)a$' DNA ta#ama# $a&' +'au *Brassica sp%+ *a!at *')'+at !a*a tae) e'"ut '#'
N.
ama
Ketea#(a#
1
Dau# +a$') !e#umu"a# *a# *'tama+"a# u=e e"$ta"$' *a# *'<.teG a
2
Dau# u=e e"$ta"$' Tee#tu" 2 -a$e $ete)a+ !e#ama+a# ")..-.m *a# '$.am') a)".+.) 3241 1G <.)um $am!e) *a# *'$e#t'-u(a$'
a
Fa$e aue.u$ 3$u!e#ata# %a#( me#(a#*u#( DNA
a
6
Fa$e .(a#'" 3*e'$ *a# !.te'# %a#( te*e#atua$' Su!e#ata# %a#( te)a+ *'!'#*a+"a# "e tau#( m'". au
a
$u!e#ata#t
Su!e#ata# %a#( te)a+ *'tama+"a# '$.!.!a#.) *'#('# 1 G <.)um $am!e) a
Te)'+at e#a#( e#a#( t'!'$ DNA Pe)et %a#( te)a+ *'tama+"a# *e#(a# 100 :u=e TE
a
u=e TE Pe)et 3me#(a#*u#( DNA
Be*a$a"a# +a$') !a"t'"um '$.)a$' DNA ta#ama# Sa&' +'au 3 Brassica sp% , )a#("a+ !etama %a#( *')a"u"a# a*a)a+ !e#(+a#5ua# mema# *a# *'#*'#( $e) !a*a *au# mu*a %a#( *'(u#a"a# I$.)a$' DNA ta#ama# '#' *'(u#a"a# *au# %a#( ma$'+ mu*a +a) '#' *'"ae#a"a# *'!u5u" *au# *a!at me#e"a# $e#%a&a !.)'-e#.) *a# !.)'$a"a'*a $e+'#((a *a!at mem!ee$a "emu#("'#a# "ee+a$')a# u#tu" me)a"u"a# '$.)a$' DNA %a#( "'ta '#('#"a# Ha) '#' *'!e"uat *e#(a# !e#%ataa# ua'*a+ 32004, !a*a a('a# ta#ama# a#%a" mem')'"' $e#%a&a !.)'-e#.) *a# !.)'$a"a'*a *a)am ".#$e#ta$' t'#((' %a#( *a!at me#(+amat !emu#'a# DNA, $e#%a&a !.)'-e#.) *a# !.)'$a"a'*a u(a *a!at mem!e#(au+' e#'me#'m $e!et' !.)'mea$e, )'(a$e, e#*.#u")ea$e e$t'"$', atau e#'m %a#( me#(a"'at"a# DNA t'*a" *'(u#a"a# *a)am !a"t'"um B'.).(' M.)e"u)e Pe#(+a#5ua# etuua# u#tu" meu$a" a'#(a# %a#( a*a !a*a $e) $e+'#((a mem!emu*a+ a+a#a+a# "'m'a )a'# %a#( a"a# *'(u#a"a# u#tu" ma$u" "e *a)am .(a#e).(a#e) $e), *a)am +a) u#tu" me#(am') DNA DNA %a#( *''$.)a$' *a' ta#ama# $e'#("a)' te".#tam'#a$' .)e+ !.)'$a"a'*a *a# meta.)'t $e"u#*e $e!et' ta#'#, !'(me#, a)"a).'* *a# a<.#.'* Se*a#("a# DNA *a' +e&a# )e'+ a#%a" me#(a#*u#( !.te'# Sa)a+ $atu "e$u)'ta# '$.)a$' DNA *a' ta#ama# t'#((' a*a)a+ !.$e$ *e$tu"$' *'#*'#( $e) u#tu" me)e!a$"a# '$' $e) Ha) '#' *'$ea"a# "ae#a ta#ama# mem')'"' *'#*'#( $e) %a#( "uat *a# $e'#("a)' !a*a eea!a e#'$ ta#ama#, ".#tam'#a$'
te$eut $u)'t *'!'$a+"a# *a' e"$ta" a$am #u")eat Pemu"aa# $e) u#tu" me#(e)ua"a# a$am #u")eat#%a !a*a !a"t'"um '#' *e#(a# 5aa me"a#'" %a'tu *'(eu$ *e#(a# t'! $te') !a*a tau#( m'". atau *a!at u(a me#((u#a"a# a+a# "'m'a %a'tu #'t.(e# 5a' u#tu" me#*e(a*a$' *a# me)aut"a# ".m!.#e# *'#*'#( $e) Pa*a $aat !.$e$ *e$tu"$' a'#(a# ta#ama# *a!at me#%ea"a# *e(a*a$' !a*a DNA *e#(a# a*a#%a a"t'<'ta$ e#'m e#*.#u")ea$e, "ae#a 'tu *'(u#a"a# u=e e"$ta"$' %a#( me#(a#*u#( $e#%a&a 200 mM T'$HC) 3!H 8, 20 mM NaC), 2 mM EDTA, *a# 0, ? SDS DNA a"a# te!'$a+ me#a*' $uatu )auta# *a# *a!at *'mu#'"a# 3*'!u'@"a$' me)a)u' *ua 5aa %a#( umum *')a"u"a# %a'tu $e#t'-u(a$' *a# e"$ta"$' "'m'a Sam!e) *'$e#t'-u(a$' *e#(a# "e5e!ata# %a#( t'#((' $e+'#((a ".m!.#e# %a#( eu"ua# )e'+ e$a atau )e'+ eat a"a# me#(e#*a! meme#tu" $e*'me# !a*a a('a# a&a+ tau#( SDS e-u#($' u#tu" me)'$'$ *'#*'#( $e) tumu+a# %a#( te*a!at *a)am )auta# $am!e) I#' *'$ea"a# "ae#a $'-at *a' SDS $ama *e#(a# $'-at *'#*'#( $e) %a#( +'*.-.'", $e+'#((a tea*' '"ata# *'a#taa "e*ua#%a *a# me#%ea"a# *'#*'#( $e) u$a" Sete)a+ *'tu#((u eea!a me#'t, $am!e) $e)a#ut#%a *''#"ua$' Mu)a*#. 32002, u=e )%$'$ '#' me#(a#*u#( EDTA %a#( e-u#($' meu$a" *'#*'#( $e) $e5aa "'m'a&' *e#(a# 5aa me#('"at '.# ma(#e$'um e-u#($' mem!eta+a#"a# '#te('ta$ $e) mau!u# a"t'@ta$ e#'m #u5)ea$e %a#( meu$a" a$am #u")eat K.t.a# 3*e'$ %a#( *'+a$')"a# *a' a"t'<'ta$ )%$'$ '#' *'e$'+"a# *e#(a# 5aa $e#t'-u(a$' a(a ".t.a# me#(um!u) *'*a$a tau#( m'". U#tu" meme$'+"a# !.te'# *a# !.)'$a"a'*a *a' )auta# *'(u#a"a# ")..-.m $e*a#("a# ".#*'$' %a#( )e'+ e"$tem, *a!at *'(u#a"a# !.te'#a$eDNA %a#( te)a+ *'e"$ta"$' *a' *a)am $e) $e)a#ut#%a !e)u *'!'$a+"a# *a' ".#tam'#a# ".m!.#e# !e#%u$u# $e) )a'##%a $e!et' !.)'$a"a'*a *a# !.te'# a(a DNA %a#( *'*a!at"a# mem')'"' "emu#'a# %a#( t'#((' Ta+a! $e)a#ut#%a %a'tu $am!e) %a#( te)a+ <.teG $e)ama *et'" *a#"emu*'a# *''#"ua$'"a# '$.)a$' !a*a $u+u 0C $e)ama 60 me#'t, +a) '#' etuua# a(a e#'m e"ea $e5aa .!t'ma) $ete)a+ *'tama+"a# u=e e"$ta"$' Kemu*'a# *')a"u"a# *'tama+"a# ")..-.m *a# '$.am') a)".+.) 3241 1G <.)um $am!e) *a# *'$e#t'-u(a$' u#tu" me#*a!at"a# $u!e#ata# Tee#tu" 2 -a$e $ete)a+ !e#ama+a# CIA *a# *'$e#t'-u(a$' %a'tu -a$e 5a' 3 $u!e#ata# *a# -a$e .(a#'" Ha) te$eut $e$ua' *e#(a# !e#%ataa# P+ama&at', 32009 me#%eut"a# a+&a $ete)a+ $e#t'-u(a$' a"a# tee#tu" 2 -a$e %a#( te!'$a+ %a"#' -a$e .(a#'" !a*a )a!'$a# a&a+ *a# -a$e au.eu$ 3a' !a*a )a!'$a# ata$ $e*a#("a# DNA *a# RNA a"a# ea*a !a*a -a$e au.eu$ $ete)a+ $e#t'-u(a$' $e*a#("a# !.te'# %a#( te*e#atua$' a"a# ea*a !a*a '#te-a$e *a# )'!'* a"a# ea*a !a*a -a$e .(a#'"
A$am #u")eat a*a)a+ m.)e"u) +'*.@)'" *a# e$'-at )aut *a)am a' D'$am!'#( 'tu, !.te'# u(a me#(a#*u#( e$'*u +'*.-.'" %a#( me#(a"'at"a# !.te'# )aut *a)am !e)aut .(a#'" Be*a$a"a# $'-at '#', te*a!at eea!a met.*e *e!.te'#'$a$' e*a$a"a# !em')'+a# !e)aut .(a#'" B'a$a#%a !e)aut .(a#'" %a#( *'(u#a"a# a*a)a+ -e#.) atau ")..-.m %a#( me#(a#*u#( 4? '$.am') a)".+.) Pe#((u#aa# ")..-.m '$.am') a)".+.) 3CIA e*a$a"a# !ee*aa# $'-at !e)aut .(a#'" K)..-.m t'*a" *a!at e5am!u *e#(a# a' *a# "emam!ua##%a u#tu" me#*e!.te'#'$a$' e*a$a"a# "emam!ua# a#ta' !.)'!e!t'*a %a#( te*e#atua$' u#tu" ma$u" atau tem.')'$a$' "e *a)am -a$e a#taa ")..-.m 7 a' K.#$e#ta$' !.te'# %a#( t'#((' !a*a -a$e a#taa te$eut *a!at me#%ea"a# !.te'# me#(a)am' !e$'!'ta$' Se*a#("a# )'!'* *a# $e#%a&a .(a#'" )a'# a"a# te!'$a+ !a*a )a!'$a# ")..-.m 3D.%)e, 1990 P.$e$ *e!.te'#'$a$' %a#( e-e"t'- e(a#tu#( !a*a e$a#%a -a$e a#taa ")..-.ma' P.$e$ '#' *a!at *')a"u"a# *e#(a# meme#tu" emu)$' *a' a' *a# ")..-.m Ha) '#' +a#%a *a!at *')a"u"a# *e#(a# !e#((..(a# atau $e#t'-u(a$' %a#( "uat "ae#a ")..-.m t'*a" *a!at e5am!u *e#(a# a' I$.am') a)".+.) e-u#($' $ea(a' emu)$'@e *a!at *'tama+"a# "e ")..-.m u#tu" mema#tu !eme#tu"a# emu)$' *a# me#'#("at"a# )ua$ !emu"aa# ")..-.ma' %a#( ma#a !.te'# a"a# me#(a)am' !e$'!'ta$' Pe#((u#aa# ")..-.m '$.am') a)".+.) '#' memu#("'#"a# u#tu" *'*a!at"a# DNA %a#( $a#(at mu#', #amu# *e#(a# u"ua# %a#( teata$ 32000070000 ! DNA "emu*'a# *''"at *a' -a$e au.eu$ *e#(a# !e$'!'ta$' '$.!.!a#.) 3Mu)a*#., 2002 Sete)a+ !.$e$ e"$ta"$', DNA %a#( *'*a!at *a!at *'!e"at"a# me)a)u' !e$'!'ta$' Pa*a umum#%a *'(u#a"a# '$.!.!a#.) *'#('# u#tu" me#(e#*a!"a#, me)e"at"a# *a# mem'$a+"a# DNA *a' )auta# Pa*a $aat !e#ama+a# '$.!.!a#.) *'#('# te)'+at e#a#(e#a#( t'!'$ DNA ta#ama# e&a#a !ut'+ %a#( me)a%a#( *' )auta# te$eut Ke*ua $e#%a&a te$eut a"a# mem!e$'!'ta$' DNA !a*a -a$e au.eu$ $e+'#((a DNA me#((um!a) meme#tu" $tu"tu @e *a# tee#tu" !e))et $ete)a+ *')a"u"a# $e#t'-u(a$' Me#uut He)*t 3200 a+&a !e$'!'ta$' e-u#($' u#tu" me#(+')a#("a# e$'*u e$'*u ")..-.m %a#( ea$a) *a' ta+a!a# e"$ta"$' Me#uut Yu&.#., 3200> !'#$'!!'#$'! !e$'!'ta$' a#taa )a'# !etama, me#uu#"a# "e)auta# a$am #u")eat *a)am a' Ha) '#' *'"ae#a"a# m.)e"u) a' %a#( !.)a me#(e)')'#(' m.)e"u) DNA *' )auta# au.eu$ Muata# *'!.)e !.$'t'- *a' a' e'#tea"$' *e#(a# muata# #e(at'- !a*a (u(u$ -.$-.*'e$te DNA I#tea"$' '#' me#'#("at"a# "e)auta# DNA *a)am a' I$.!.!a#.) *a!at e5am!u *e#(a# a', #amu# "ua#( !.)a *'a#*'#("a# a' M.)e"u) '$.!.!a#.) t'*a" *a!at e'#tea"$' *e#(a# (u(u$ !.)a *a' a$am #u")eat $e+'#((a '$.!.!a#.) a*a)a+ !e)aut %a#( )ema+ a(' a$am #u")eatQ "e*ua,
!e#ama+a# '$.!.!a#.) a"a# me#(+')a#("a# m.)e"u) a' *a)am )auta# DNA $e+'#((a DNA a"a# te!e$'!'ta$'Q "et'(a, !e#((u#aa# '$.!.!a#.) *'#('# a"a# me#uu#"a# a"t'<'ta$ m.)e"u) a' $e+'#((a memu*a+"a# !e$'!'ta$' DNA Pa*a ta+a!a# !e$'!'ta$' '#', DNA %a#( te!e$'!'ta$' a"a# te!'$a+ *a' e$'*ue$'*u RNA *a# !.te'# %a#( ma$'+ te$'$a Re$'*u te$eut u(a me#(a)am' ".a(u)a$'#amu# t'*a" meme#tu" $tu"tu @e *a# ea*a *a)am e#tu" !e$'!'tat (a#u)aPa*a $aat eta#.) atau '$.!.!a#.) *'ua#( *a# !e))et *'"e'#(a#('#"a# *a)am tau#(, ma"a !e))et %a#( te$'$a *a)am tau#( a*a)a+ DNA !e"at P.$e$ !e$'!'ta$' "ema)' *e#(a# '$.!.!a#.) $ee)um !e))et *'"e'#(a#('#"a# *a!at me#'#("at"a# *eaat "emu#'a# DNA %a#( *''$.)a$' 3Fae), 2004 Ta+a! $e)a#ut#%a %a'tu tau#( *'".5." *a# *''#"ua$' !a*a $u+u 200C $e)ama 1 me#'t Ha) '#' *')a"u"a# u#tu" .!t'ma)'$a$' "ea '$.!.!a#.) Kemu*'a# *'$e#t'-u(a$' *e#(a# "e5e!ata# 000 !m G ( $e)ama 1 me#'t P.$e$ $e#t'-u(a$' u)a#(a# *')a"u"a# u#tu" me#*a!at"a# !e))et DNA Ta+a! $e)a#ut#%a %a'tu tau#( *' $e#t'-u(a$' "ema)' !a*a "e5e!ata# 000 !m $e)ama 1 me#'t Ha$') %a#( *'*a!at %a'tu eu!a $u!e#ata# %a#( e#'#( *a# !e)et 3e#*a!a# DNA mu#' %a#( e&a#a !ut'+ te*a!at !a*a *a$a tau#( Kemu*'a# $u!e#ata# te$eut *'ua#( "ae#a DNA ea*a !a*a a('a# #ata# Me#uut D.#ata 32008 DNA mu#' %a#( *'+a$')"a# a*a)a+ DNA %a#( teea$ *a' 5am!ua# mate'a) *a# ".m!.#e# '#ta5e)u)e %a#( me#(a#*u#( )auta# ".m!)e"$ eu!a RNA, !.te'#, )ema" *a# "a.+'*at Sete)a+ 'tu*'tama+"a# 100 : u=e TE "e *a)am tau#( %a#( e'$' !e)et *a# "emu*'a# *'$'m!a# *' *a)am -eee *e#(a# $u+u $e"'ta 20C A*'a#a, 32009 me#%ata"a# a+&a u=e TE *a# !e#%'m!a#a# $u+u !a*a 20C etuua# a(a $am!e) DNA %a#( te)a+ *'e"$ta"$' *a!at *'$'m!a# +'#((a &a"tu em'#((um'#((u P+ama&at', 32009 u(a me#e)a$"a# a+&a !e)auta# "ema)' *e#(a# u=e TE u(a *a!at mem'$a+"a# a#taa RNA %a#( mem!u#%a' eat m.)e"u) )e'+ e#*a+ *'a#*'#("a# DNA $e+'#((a DNA %a#( *'*a!at"a# t'*a" te".#tam'#a$' .)e+ RNA *a# DNA $a#(at $ta') "et'"a *'$'m!a# *a)am "ea*aa# te!e$'!'ta$' !a*a $u+u 20C V"
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DAFTAR PUSTAKA
A*'a#a, D" 2009 Te"#'" '$.)a$' DNA (e#.m ta#ama# !a!a%a *a# eu" *e#(a# me#((u#a"a# m.*'@"a$' u=e CTAB% Buletin Teknik )ertanian 14(1): 121
D.#ata 2008 Komunikasi pribadi% Ciri3ciri D#A murni dan penyebab keberhasilan serta kegagalan dalam )C4 dan elektroforesis% E)a#((a ;a"ata D.%)e ;; a#* D.%)e ;: 1990 I$.)at'.# .- !)a#t DNA -.m -e$+ t'$$ue F.5u$ M.$5.& 1231161 Fae), RE 2004 4#A ethodologies0 A 5aboratory $uide for "solation and Characteri6ation% Third Edition% E)$e<'e A5a*em'$ Pe$$ :.#*.# Ha) 9680 He)*t, H 200 )lant Biochemistry% Third Edition% E)$e<'e A5a*em'5 Pe$$ Ca)'-.#'a Ha) 491D.%)e ;;, D.%)e ;: 1990 I$.)at'.# .- !)a#t DNA -.m -e$+ t'$$ue 7ocus 12 16 1 Mu)a*#. 32002 Se!uta Te"#.).(' Re"a%a$a e#et'"a Pu$ata"a 'au$a+a Mu*a B.(. P+ama&at', M 2009 O!t'ma)'$a$' e"$ta"$' DNA *a# PCRRAPD !a*a (e<'))ea $!! 3P.tea5eae 8urnal Biologi 13(1): 121 R'e'. RA, :.8 Yu&.#., T'&'.&. 200> B'.).(' M.)e"u)e ;a"ata Pe#e't E)a#((a
Lukisan Karya 'amis, 20 &aret 2014
Laporan Genetika Isolasi DNA LAPORAN PRAKTIKUM
GENETIKA PERCOBAAN II
ISOLASI DNA NAMA NIM KELOMPOK HARI/TGL" PERCOBAAN ASISTEN
: NUR AFIAH SULAIMAN : H41113704 : V (LIMA) B : KAMIS/ 13 MARET 2014 : RISK NURHIKMAANI
LABORATORIUM GENETIKA !URUSAN BIOLOGI FAKULTAS MATEMATIKA DAN ILMU PENGETAHUAN ALAM UNIVERSITAS HASANUDDIN MAKASSAR 2014 BAB I
PENDAHULUAN I"1 L#$#% B&'##*
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etem!at *' :a.at.'um B'.).(' Da$a, Fa"u)ta$
Matemat'"a *a# I)mu Pe#(eta+ua# A)am, U#'
BAB II
TIN!AUAN PUSTAKA
Stu*' me#(e#a' e"$'$te#$' a$am #u")eat !etama "a)' *')a"u"a# .)e+ F'e*'5+ M'e$5+e *a' ;ema# %a#( me#('$.)a$' '#t' *a' $e) *aa+ !ut'+ !a*a ta+u# 1>9 M'e$5+e me#emu"a# a+&a *' *a)am '#t' $e) te$eut te*a!at $e#%a&a %a#( me#(a#*u#( -.$-at %a#( "emu*'a# *'#ama"a# #u")e'# Se)a#ut#%a !a*a a"+' aa* "e19 te)a+ e+a$') *')a"u"a# !em'$a+a# a#taa DNA 3 deoxyribonucleic acid *a# RNA 3 ribonucleic acid *a' !.te'#!.te'# %a#( me)e"at"a# m.)e"u) a$am
#u")eat te$eut !a*a $e) Pa*a a&a) ta+u# 1960a#, P :e
%a#( $ama *a# te(a" )uu$ te+a*a! a"$'$ u#ta'a# DNA U#ta'a# DNA mem!u#%a' *ua )e"u"a# 3groo1e e"$te#a) %a'tu )e"u"a# e$a 3 ma(or groo1e *a# )e"u"a# "e5') 3minor groo1e Ke*ua )e"u"a# te$eut mem!u#%a' !ea#a# $ea(a' tem!at me)e"at#%a m.)e"u) !.te'# tete#tu 3Yu&.#., 200 U"ua# m.)e"u) DNA e
DNA
a*a
2, %a'tu $e#t'-u(a$'
*a#
!e$'!'ta$' P'#$'! utama
$e#t'-u(a$' a*a)a+ mem'$a+"a# $u$ta#$' e*a$a"a# eat e#'$ m.)e"u) *e#(a# 5aa meme'"a# (a%a $e#t'-u(a) $e+'#((a $u$ta#$' %a#( )e'+ eat a"a#ea*a *' *a$a, $e*a#("a# $u$ta#$' %a#( )e'+ '#(a# a"a# te)eta" *' ata$ Te"#'" $e#t'-u(a$' te$eut *')a"u"a# *' *a)am $eua+ me$'# %a#( e#ama me$'# $e#t'-u(a$' *e#(a# "e5e!ata# %a#( e
*e#(a# ua+ e"a*a a' e#*a Sema"'# t'#((' "a*a a' ma"a $e) %a#( te)aut *' *a)am e"$ta" a"a# $ema"'# $e*'"'t, $e+'#((a DNA %a#( te!e$'!'ta$' u(a a"a# $e*'"'t 3A(u$ *a# Sa-aae#a#, 2014 P.$e$ '$.)a$' DNA *'a&a)' *e#(a# !.$e$ e"$ta"$' DNA Ha) '#' etuua# u#tu" mem'$a+"a# DNA *e#(a# !at'"e) )a'# %a#( t'*a" *''#('#"a# P.$e$ '#' +au$ *')a"u"a# *e#(a# +at'+at', $e+'#((a t'*a" me#%ea"a# "eu$a"a# !a*a *'#*'#( $e), mema# $e) mema# !)a$ma *a# mema# '#t' a'" *e#(a# 5aa me"a#'" mau!u# $e5aa "'m'a&' Caa me"a#'" '$a *')a"u"a# *e#(a# !em)e#*ea# atau !e#((eu$ me#((u#a"a# m.ta *a# !'$t') Se*a#("a# $e5aa "'m'a&' *a!at *e#(a# !eme'a# %a#( *a!at meu$a" mema# $e) *a# mema# '#t', $a)a+ $atu#%a a*a)a+ *ete(e# 3A(u$ *a# Sa-aae#a#, 2014 Se)a'#
'tu,
*e#(a# me#ama+"a#
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$ee#'$ *etee# *a!at *'(u#a"a#
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+a#%a m.)e"u) #u")e.t'*a 3DNA *a# RNA U#tu" me#(+')a#("a#
!.te'# *a' )auta#,
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!+e#.) 3me#('"at !.te'# *a# $ea('a# "e5') RNA *a#5+)..-.m 3meme$'+"a# !.te'# *a# !.)'$a"a'*a *a' )auta# Eta#.) e-u#($' u#tu" meme"at"a#, mem'$a+"a# DNA *a' )auta# *a# me#(e#*a!"a# DNA3Mu)a*#., 200 2 Pe#ama+a# *ete(e# *a)am '$.)a$' DNA *a!at *')a"u"a# "ae#a *ete(e# *a!at me#%ea"a# ua"#%a mema# $e), me)a)u' '"ata# %a#( *'e#tu" me)a)u'
$'$' +'*.-.'" *ete(e# *e#(a# !.te'# *a# )ema" !a*a mema# meme#tu" $e#%a&a )'!'* !.te'#*ete(e# ".m!)e"$ Se#%a&a te$eut *a!at tee#tu" "ae#a !.te'# *a# )'!'* mem')'"' uu#( +'*.@)'" *a# +'*.-.'", *em'"'a# u(a *e#(a# *ete(e# $e+'#((a *a!at meme#tu" $uatu '"ata# "'m'a Kemu*'a# !e#ama+a# (aam 3NaC) e-u#($' u#tu" me)aut"a# DNA *a# $ete)a+ 'tu *'e'"a# et+a#.) *'#('# 9? u#tu" me#(um!u)"a#/me#((um!a)"a# DNA 3A(u$ *a# Sa-aae#a#, 2014 E)e"t.-.e$'$ DNA meu!a"a# te"#'" u#tu" mem'$a+"a# $am!e) DNA e*a$a"a# ata$ u"ua# 3eat m.)e"u) *a# $tu"tu @$'" m.)e"u)#%a e) %a#( 'a$a *'(u#a"a# a#taa )a'# a(a.$a E)e"t.-.e$'$ (e) a(a.$a *a!at *')a"u"a# u#tu" mem'$a+"a# $am!e) DNA *e#(a# u"ua# *a' eea!a atu$ +'#((a 20000 !a$a#( a$a 3! 3D&'*.$e!ut., 199> M.)e"u) DNA emuata# #e(at'- $e+'#((a *' *a)am me*a# )'$t'" a"a# em'(a$' me)a)u' mat'"$ (e) me#uu "utu !.$'t'- 3a#.*e Ma"'# e$a u"ua# m.)e"u)#%a, ma"'# e#*a+ )au m'(a$'#%a Beat m.)e"u) $uatu -a(me# DNA *a!at *'!e"'a"a# *e#(a# mema#*'#("a# )au m'(a$'#%a *e#(a# )au m'(a$' -a(me# -a(me# m.)e"u) DNA $ta#*a 3DNA ma"e %a#( te)a+ *'"eta+u' u"ua##%a V'$u)'$a$' DNA $e)a#ut#%a *')a"u"a# *' a&a+ !a!aa# $'#a u)ta<'.)et $ete)a+ te)e'+ *a+u)u (e) *a)am !emuata##%a *'tama+"a# )auta# et'*'um .m'* Caa )a'# u#tu" me)'+at <'$ua)'$a$' DNA a*a)a+ (e) *'e#*am *' *a)am )auta# et'*'um .m'* $ee)um *'!a!a"a# *' ata$ $'#a u)ta<'.)et 3D&'*.$e!ut., 199> ;um)a+ DNA *'5em'#"a# eat m.)e"u) u"a# .)e+ <.)ume U#tu" me#(eta+u' um)a+ DNA, ma"a DNA +a$') '$.)a$' +au$ *'a#a)'$'$ *e#(a# $!e"t.-.t.mete UV*e #(a# !a#a#( (e).ma#( 2 #m 320 #m Kua)'ta$ DNA %a#( e+uu#(a# *e#(a# a$.a#$'
"emu#'a# te+a*a! ".#tam'#a# !.te'# *a!at *')'+at *a' !ea#*'#(a# $u$!e#$' DNA !a*a !a#a#( (e).ma#(
3Su+a$.#. *a# '*%a$tut', 200 BAB III
20 #m te+a*a! 2>0 #m
METODE PERCOBAAN
III"1 A'#$
A)at %a#( *'(u#a"a# *a)am !e5.aa# '#' a*a)a+ !'$au, me$'# )e#*e, t'ma#(a#, (e)a$ aua, !e#(a*u", !e#%a'#( 3t'$$u / "a!a$ / "eta$ $a'#(, $!atu)a, tau#( ea"$', !'!et tete$, *a# me$'# <.teG III"2 B##
Ba+a#
%a#( *'(u#a"a#
a*a)a+ ua+
t.mat Solanum lypersium, ua+
!e!a%aCarica papaya, *ete(e# Su- uu", *ete(e# R'#$. 5a', $au# Bu5eam, aua*e$, (aam *a!u, *a# eta#.) 9? *'#('# III"3 P%.5&+% K&%#
A*a!u# )a#("a+)a#("a+ "ea %a#( *')a"u"a# *a)am !e5.aa# '#' $ea(a' e'"ut 1
Me)aut"a# *ete(e# 3Su-, R'#$. *a# Bu5eam "e *a)am 0 m: aua*e$ *'a*u" !e)a# $e)ama 1 me#'t
2
Me#(am') 100 (m *a('#( ua+ *'tama+ 100 m: aua*e$ *'ma$u""a# "e *a)am me$'# )e#*e, "emu*'a# *')e#*e $e)ama 40 *et'"
6
Me#5am!u"a# 4 m: ma$'#(ma$'#( )auta# $au# *e#(a# ma$'#(ma$'#( 4 m: u$ ua+
4
Me#ama+"a# 1 $!atu)a (aam *a!u "emu*'a# *'a*u" $e)ama 10 me#'t $am!a' *'!e.)e+ 5am!ua# %a#( +.m.(e# $eta *'<.teG
Me#%a'#( 5am!ua# %a#( *'+a$')"a# $ee)um#%a $ea#%a" *ua "a)' !e#%a'#(a#
m: +a$') !e#%a'#(a# te$eut "emu*'a# *'ma$u""a# "e *a)am tau#( ea"$' *a# me#ama+"a# m: eta#.) 9 ? *'#('#
8
Me#(amat' !.$e$ t'mu)#%a DNA, me)'!ut' &a"tu %a#( *'!e)u"a#, &a#a, $eta a#%a" $e*'"'t#%a DNA %a#( tee#tu"
W
BAB IV
HASIL DAN PEMBAHASAN
IV"1 H#56' P&%-.##
IV11 ama +a$') Pe5.aa# ;e#'$ ua+ T.mat
See)um Dete(e# Buu"
Dete(e# 5a'
Dete(e# "'m
Pe!a%a
Dete(e# Buu"
Dete(e# 5a'
Se$u*a+
Dete(e# "'m
IV"1"2 T#&' P&*##$#
N.
;e#'$ Bua+
Pe)a"ua#
Ha$') Pe#(amata# a#a
1
T.mat Solanumlycopersiu m
;um)a+
a"tu
Dete(e#
Be#'#(,
a*a :amat
Buu"
e#*a!a#
Dete(e#
Be#'#(
Se*a#(
Put'+ Be#'#(
:e'+
Ca' Dete(e# K'm 2
Pe!a%a Carica papaya
Ce!at
Dete(e#
Put'+
:e'+
Buu"
Ke"u#'#(
Ce!at
"u#'#(a# Dete(e#
Put'+ "eu+
:amat
Put'+ $u$u
Se*a#(
Ca' Dete(e# K'm
IV"2 P&##5#
Pa"t'"um '$.)a$' DNA '#' *')a"u"a# etuua# u#tu" me#(eta+u' !e#(au+ ma5am ua+ *a# e#'$ *etee# te+a*a! "ua)'ta$ DNA %a#( *'+a$')"a# *a)am !.$e$ '$.)a$' Se+'#((a *a)am !a"t'"um '#' *'uat
*ete(e# *a# e#'$ ua+ a(a *a!at me#(eta+u' ua+ *a# e#'$ *ete(e# ma#a %a#( me#(+a$')"a# DNA !a)'#( a(u$ *a# a'" ;e#'$ ua+ %a#( *'(u#a"a# a*a)a+ t.mat Solanum lycopersicum $ea(a' 5.#t.+ ua+ %a#( me#(a#*u#( a#%a" a' *a# !e!a%a Carica papaya $ea(a' ua+ %a#( mem')'"' "a#*u#(a# a' t'*a" te)a)u a#%a" Se*a#("a# u#tu" *ete(e# %a#( *'!a"a' a*a)a+ *ete(e# uu" me" Su-, *ete(e# 5a' me" R'#$., *a# *ete(e# "'m me" BuCeam Pa*a *a$a#%a !'#$'! '$.)a$' DNA a*a t'(a %a'tu !e)'$'$a# $e), e"$ta"$' *a# !emu#'a# Pa*a
!a"t'"um
"a)'
'#'
%a#(
!etamatama
*')a"u"a#
'a)a+
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ua+ *'tama+"a#
*e#(a# )auta#
*ete(e# Pe#ama+a#
)auta# *ete(e# e-u#($' u#tu" me)'$'$ *'#*'#( $e) tumu+a# %a#( te*a!at *a)am )auta# $am!e) I#' *'$ea"a# "ae#a $'-at *a' *ete(e# $ama *e#(a# $'-at *'#*'#( $e) %a#( +'*.-.'", $e+'#((a tea*' '"ata# *'a#taa "e*ua#%a *a# me#%ea"a# *'#*'#( $e) u$a" :a)u *'tama+"a# (aam *a!u *a# *'a*u" Pe#('$.)a$'a# DNA me#((u#a"a# (aam *a!u *e#(a# tuua# u#tu" meme"at"a# DNA Ha) '#' *a!at tea*' "ae#a '.# Na %a#( *'"a#*u#( .)e+ (aam mam!u meme#tu" '"ata# *e#(a# "utu #e(at'
)auta# *'$a'#( *e#(a# "eta$
$a'#( $ea#%a" *ua
"a)'
!e#%a'#(a#u#tu" mem'$a+"a# $eat$eat %a#( "a$a *e#(a# %a#( +a)u$, $e+'#((a *'*a!at"a# $am!e) eu!a 5a'a# %a#( t'*a" te)a)u "e#ta) Ha$') !e#%a'#(a# te$eut *'ma$u""a# "e *a)am tau#( ea"$' *a# *'tama+ eta#.) 9 ?
*'#('# %a#( e-u#($' mema#tu !.$e$ !e#(e#*a!a# te+a*a!
.(a#e).(a#e) %a#( $u*a+ "e)ua *a' $e) atau mem'$a+"a# a('a#a('a# %a#(
teua' te$eut e*a$a"a# eat m.)e"u) Et+a#.) e!ea# *a)am !e#(um!u)a# *a# !e#((um!a)a# DNA "ae#a $'-at#%a %a#( *'#('# Kemu*'a# $ete)a+ *'amat' !.$e$ t'mu)#%a DNA, me)'!ut' &a"tu %a#( *'!e)u"a#, &a#a, $eta a#%a" $e*'"'t#%a DNA %a#( tee#tu" Ma"a +a$') %a#( *'*a!at"a# %a'tu )auta# %a#( !a)'#( 5e!at meme#tu" (um!a)a# DNA a*a)a+ t.matBu"'m *a# t.matSu- Ha) '#' et.)a" e)a"a#( *e#(a# te.' %a#( me#%ata"a# a+&a ua+ !e!a%a a"a# )e'+ a#%a" me#(+a$')"a# (um!a)a# DNA "ae#a mem')'"' "a#*u#(a# a' %a#( )e'+ $e*'"'t *a'!a*a t.mat $e+'#((a DNA %a#( te!e$'!'ta$' )e'+ a#%a" !a*a ua+ !e!a%a Ke#%ataa# '#' u(a t'*a" $ea)a# *e#(a# a#((a!a# *ete(e# %a#( !a)'#( a'" *'(u#a"a# u#tu" '$.)a$' DNA a*a)a+ *ete(e# uu", "emu*'a# *ete(e# 5a', )a)u *ete(e# "'m Da' +a$') te$eut ma"a %a#( *a!at *'$'m!u)"a# a+&a t.mat *a# BuCeam !a)'#( a#%a" me#(+a$')"a# DNA Da# *a' "et'(a e#'$ *etee# %a#( *'(u#a"a#, %a#( !a)'#( $e*'"'t me#(+a$')"a# DNA a*a)a+ *etee# R'#$. :auta# *e#(a# *etee# uu" Su- *a# Bu "'m me#(+a$')"a# DNA *e#(a# um)a+ %a#( 5u"u! a#%a" Namu# "et'(a e#'$ *etee# te$eut t'*a" *a!at *'a#*'#("a# ma#a %a#( )e'+ a'" *a)am me#('$.)a$' DNA "ae#a !a*a $et'a! !e)a"ua# me#u#u""a# +a$') %a#( e)at'- $ama Kemu#("'#a# te*a!at "e$a)a+a# !a*a $aat *')a"u"a# !em)e#*ea# ua+ !e!a%a %a#( te)a)u )ama *a# te)a)u +a)u$ $e+'#((a DNA %a#( *'*a)am#%a te5a'"5a'" +a#5u *a# me#%ea"a# "e(a(a)a# !a*a $aat !a*a '$.)a$' DNA Se)a'# 'tu !a*a $aat !.$e$ !e#%a'#(a# *a# !e#ama+a# et+a#.) tea*' "e+a'$a# a+a# *a# "ae#a#%a te*a!at $ea('a# !e5.aa# %a#( e)um tu#ta$ *a# me#((u#a"a# a+a# !e#((a#t' %a#( teata$ "a!a$'ta$#%a *'a#*'#("a# a+a# %a#( $eme$t'#%a *'(u#a"a# *a)am !a"t'"um
BAB V
PENUTUP V"1 K&56+'#
A*a!u# "e$'m!u)a# *a' !a"t'"um '#' a*a)a+ 1
U#tu" me)a"u"a# '$.)a$' DNA met.*e atau te"#'" %a#( *'(u#a"a# !a*a *a$a#%a a*a t'(a %a'tu !e)'$'$a# $e), e"$ta"$', *a# !emu#'a# Pe)'$'$a# *a!at *')a"u"a# *e#(a# 5aa me"a#'" mau!u# "'m'a, $a)a+ $atu#%a *e#(a# 5aa *')e#*e *a# *'5am!u )auta# *ete(e# E"$ta"$' *a!at *')a"u"a# *e#(a# !e#ama+a# (aam *a!u 3NaC) *a# !emu#'a# $a)a+ $atu met.*e#%a a*a)a+ *e#(a# meme'"a# )auta# et+a#.) *'#('# 9?
2 ;e#'$ *ete(e# *a# ua+ %a#( *'!a"a' u#tu" me)a"u"a# '$.)a$' DNA $a#(at e!e#(au+ !a*a DNA %a#( *'+a$')"a# V"2 S#%#
A*a!u# $aa# u#tu" !a"t'"um '#' $ea'"#%a a)at *a# a+a# )a.at.'um *'tama+
u#tu"
me#5e(a+
tea*'#%a
"e"ua#(a#
a+a#
%a#(
*a!at
me#(a"'at"a# "ea$a)a+a# *a# "ua#( a"uat#%a +a$') %a#( *'*a!at"a# !a*a !a"t'"um '#'
