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Lampiran 1. Komposisi pereaksi dan bufer untuk analisis DNA klon kelapa sawit Larutan

Bufer ekstrak Orozco-Castillo et al.

(1 994) Larutan TE Larutan TAE 50X Bufer amplifikasi 10X Loading bufer Komposisi 10% CTAB 5 M NaCl 0,2 % merkaptoetanol 0,5 M EDTA pH 8 1 M Tris-HC1 pH 8 1 rnl Tris-HC1 pH 8 0,5 M EDTA pH 8 24,2 g Tris-base

5,7 ml asam asetat glacial 10 ml EDTA 0,5 M pH 8

air bebas ion hingga 100 ml

5 ml Tris-HC1 pH 8 10 mM NaCl 0,l mM EDTA 0,2 1 % triton 2,5% bromofenol blue 40% sukrosa

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52

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53 Lampiran 3. Nilai komponen utama dari pita-pita hasil amplifikasi dengan 15 primer

acak yang berperan membedakan 6 klon kelapa sawit

No. Primer Pita ke Nilai Komponen Utama

1 2 3 4 5 6 1 OPC-08 3 0.108 0.090 -0.072 0.071 -0.072 -0.083 2 4 -0.207 0.100 0.123 0.061 0.038 -0.053 3 6 0.243 -0.081 0.004 0.040 -0.068 -0.008 4 9 -0.193 0.106 -0.001 -0.104 0.106 0.083 5 OPC-09 2 -0.012 0.043 0.261 -0.059 -0.008 -0.004 6 4 0.152 -0.036 0.079 -0.145 -0.070 0.190 7 7 -0.021 0.084 -0.130 0.137 -0.091 -0.102 8 8 -0.023 -0.033 0.148 0.041 0.085 0.046 9 OPB-05 1 0.158 0.116 -0.070 0.008 -0.034 -0.007 10 2 0.060 0.065 -0.010 -0.041 0.154 0.051 11 3 0.028 0.211 0.144 -0.045 0.024 -0.056 12 4 0.046 -0.190 0.025 0.114 0.044 0.002 13 5 0.028 0.211 0.144 -0.045 0.024 -0.056 14 6 -0.022 -0.077 -0.060 0.029 -0.085 -0.206 15 8 -0.074 -0.021 -0.168 -0.068 -0.068 0.054 16 OPB-06 1 -0,011 0.184 -0.043 -0.072 -0.070 0.008 17 2 -0.075 0.063 -0.096 -0.165 -0.300 0.001 18 3 0.046 -0.190 0.025 0.114 0.044 0.002 19 4 0.158 0.116 -0.070 0.008 -0.034 -0.007 20 5 0.168 0.072 0.028 -0.140 -0.088 0.036 2 1 6 0.047 -0.198 0.031 0.065 -0.132 -0.007 22 OPA-02 1 0.016 -0.076 -0.018 -0.004 0.013 -0.200 23 2 0.000 -0.166 -0.146 -0.046 0.097 -0.013 24 OPA-04 3 0.036 -0.147 -0.020 0.055 0.067 -0.074 25 4 0.045 0.126 -0.098 -0.011 -0.168 0.125 26 5 -0.066 0.056 -0.165 0.162 0.041 0.240 27 6 0.116 -0.037 -0.150 -0.001 -0.060 0.046 28 OPN-16 3 0.043 0.123 -0.012 0.136 0.236 -0.128 29 6 0.000 -0.094 0.052 0.164 -0.036 0.235 30 8 0.036 -0.147 -0.020 0.055 0.067 -0.074 31 OPN-18 2 -0.085 0.197 -0.073 -0.032 0.034 0.001 32 4 -0.054 -0.054 0.032 0.000 -0.003 -0.205 33 5 -0.016 0.076 0.018 0.004 -0.013 0.200 34 SCIO-19 1 0.224 0.072 -0.051 -0.055 0.018 -0.234 35 4 -0.071 0.158 -0.103 0.136 0.099 0.010 36 5 0.076 0.076 -0.066 0.087 0.153 -0.053 37 7 -0.071 -0.013 -0.108 -0.142 0.056 0.051 38 8 0.076 0.113 -0.115 0.153 0.129 -0.099 39 11 -0.085 0.197 -0.073 -0.032 0.034 0.001 40 14 0.100 0.043 -0.070 0.171 0.062 0.131 41 SCIO-20 1 -0.022 0.049 0.091 0.005 0.224 -0.054 42 2 0.000 0.166 0.146 0.046 -0.097 0.013 43 5 -0.036 0.147 0.020 -0.055 -0.067 0.074 44 6 0.088 0.018 0.051 -0.145 -0.040 0.073 45 OPC-02 2 -0.150 0.045 0.014 0.097 -0.213 0.011 46 3 -0.016 0.076 0.018 0.004 -0.013 0.200

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54

Lanjutan

Akar ciri 3.6517 3.4802 1.6209 1.3788 1.0658 0.8657

proporsi 22.80% 21.70% 10.10% 8.60% 6.70% 5.40%

kumulatif 22.80% 44.50% 54.60% 63.30% 69.90% 75.30% Keterangan : Nilai komponen utama pita-pita RAPD yang berperan ditandai dengan angka tebal

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Lampiran 4. Jenis primer, total pita DNA enam Klon Kelapa sawit yang diamplifikasi menggunakan 15 primer acak. No 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Primer OPC-08 OPC-09 OPB-05 OPB-06 OPA-02 OPA-04 OPN-16 OPN-18 SC10-19 SC10-20 OPC-02 OPG07 OPH-18 OPW-19 OPE-14 -- - TOTAl RATA- RATA PITA Total pita 11 8 8 6 6 6 8 5 14 6 6 8 6 7 10 115 8

Klon Kelapa sawit dan Jumlah pita

MK152 J 4 1 1 2 5 2 3 1 3 1 3 5 5 2 7 45 MX152 ABN 5 2 2 2 4 3 4 2 3 0 5 6 5 3 6 52 MK152 N 5 3 2 2 3 4 5 1 1 3 4 4 5 3 5 50 MK209 J 5 3 3 4 4 2 3 1 4 4 4 5 5 2 6 55 MK209 ABN 8 3 4 4 4 2 3 2 5 3 4 6 5 2 6 61 MK209 N 7 2 4 3 4 4 3 3 9 3 4 5 5 3 5 64 MK212 J 7 2 4 3 4 4 3 3 10 3 4 4 5 4 5 67 a 2 1 2 ABN 9 3 5 3 4 3 3 3 7 4 4 4 5 3 6 66 MK212 N 8 3 4 1 4 4 3 3 9 4 4 3 5 4 5 64 MX203 J 7 3 3 3 3 0 3 2 4 5 4 2 2 1 4 44 MK203 ABN 7 1 3 3 3 4 2 3 4 3 4 0 6 1 4 46 MK203 N 6 1 3 3 3 3 2 3 4 4 5 5 6 2 4 52 MK176 ABN 6 0 3 3 3 4 3 2 5 4 5 5 5 2 4 53 MK176 N 6 5 3 3 3 2 0 2 4 4 5 0 5 1 4 46 M K l M ABN 4 0 1 4 4 4 0 2 4 2 4 0 5 5 8 45 M K l M N 6 3 2 3 3 4 0 2 4 3 5 6 5 2 4 52

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Lampiran 5. Jenis primer, total pita polimorfik enam Klon Kelapa sawit yang diamplifikasi menggunakan 15 primer acak. No 1. 2. 3. 4. 5. 6. 7. 8. 9 10. 11. 12. 13. 14. 15. Primer OPC08 OPC-09 OPB-05 OPB-06 OPA-02 OPA-04 OPN-16 OPN-18 SC10-19 SC10-20 0 x 4 2 OPC-07 OPH-18 OPW-19 OPE-14 TOTAL Persentase (%) Total pita poli morfik 8 8 8 6 3 6 8 5 13 6 3 8 5 7 6 100 87

Klon kelapa sawit dan Jumlah pita polimorfik

MK152 J 1 1 1 2 2 2 3 1 2 1 0 5 4 2 3 30 666 MK152 ABN 2 2 2 2 1 3 4 2 2 0 2 6 4 3 2 37 71 MK152 N 2 3 2 2 0 4 5 1 0 3 1 4 4 3 1 35 70 MK209 J 2 3 3 4 1 2 3 1 3 4 1 5 4 2 3 41 75 MK209 ABN 5 3 4 4 1 2 3 2 4 3 1 6 4 2 2 46 75 MK209 N 4 2 4 3 1 4 3 3 8 3 1 5 4 3 1 49 76,6 MK212 I 4 2 4 3 1 4 3 3 9 3 1 4 4 4 1 50 76,9 MK212 ABN 6 3 5 3 1 3 3 2 6 4 1 4 4 3 2 51 77,3 MK212 N 5 3 4 1 1 4 3 3 8 4 1 3 4 4 1 49 76,6 MK203 J 4 3 3 1 0 0 3 3 3 5 1 2 1 1 0 25 65,9 a 2 0 3 ABN 4 1 3 1 0 4 2 3 3 3 1 0 5 1 0 31 67 MK203 N 3 1 3 1 0 3 2 3 2 4 2 5 5 2 0 36 69 a 1 7 6 ABN 3 0 3 2 0 4 3 2 4 4 2 5 5 2 0 39 74 a 1 7 6 N 3 5 3 2 0 2 0 2 3 4 2 0 4 1 0 31 67,4 MK104 ABN 1 0 1 2 1 4 0 2 3 2 2 0 4 5 4 27 68.8 MK104 N 3 3 2 3 0 4 0 2 3 3 2 6 4 2 0 37 71

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Larnpiran 6. Jenis, susunan oligonukleotida dan kandungan GIC 3 1 primer acak RAPD yang diseleksi

No 1 2 3 4 5 6 7 8 9 1 0 11 1 2 1 3 1 4 1 5 1 6 1 7 1 8 1 9 2 0 2 1 2 2 2 3 24 2 5 2 6 27 28 2 9 3 0 3 1 Primer OPA-02 OPA-04 O P B - 0 5 O P B - 0 6 O P C - 0 1 OPC-02 O P C - 0 3 OPC-04 O P C - 0 5 O P C - 0 6 OPC-07 O P C - 0 8 OPC-09 O P C - 1 0 OPA-07 O P E - 1 4 OPH-18 OPN-10 OPN-16 OPN-18 OPW-15 OPW-19 S C 1 0 - 4 8 S C 1 0 - 5 6 S C 1 0 - 5 7 S C 1 0 - 7 1 S C 1 0 - 7 6 S C 1 0 - 8 3 S C 1 0 - 9 1 S C 1 0 - 1 9 S C 1 0 - 2 0 Susunan nukleotida 5'

.

... 3' TGC CGA GCTG AAT CGG GCTG TGC CTC GCCC TGC TCT GCCC TTC GAG CCAG GTG AGG CGTC GGG GGT CTTT CCG CAT CTAC GAT GAC CGCC GAA CGG ACTC GTC CCG ACGA TGG ACC GGTG CTC ACC GTCC TGT CTG GGTG GAA ACG GGTG TGC GGC TGAG GAA TCG GCCA ACA ACT GGGG AAG CGA CCTG GGT GAG GTCA ACA CCG GAAC CAA AGC GCTC CTG GTA TGCG CCA GCG TCTA GCT GGA AGCT CTG GCG TAGT CGC AGA CTTG AAT GAC GGGG CTC GAC TAGG CGT CCG TCAG ACT CGT AGCC Kandungan G / C ( 8 ) 7 0 6 0 7 0 7 0 6 0 7 0 6 0 6 0 7 0 6 0 7 0 7 0 7 0 6 0 6 0 7 0 6 0 6 0 6 0 6 0 6 0 6 0 6 0 6 0 6 0 '7 0 6 0 6 0 6 0 6 0 7 0

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Lampiran 7. Profil pita DNA seleksi primer

Keterangan : Pita DNA ke enam klon kelapa sawit hasil amplifikasi menggunakan primer

(a)OPB-05; @)SC10-20; (c)OPA-04; Lajur (M) 1 kb DNA Ladder, l)I~lK152~, 2)MKl 52,bn, 3)MK152,, 4)MK209j, 5)MK2Ogah, 6)MK209,, 7)MK2 1 2j, 8)MK212abn, 9)MK212,, I O)MK203j, 1 l)MK203abn, 1 2)MK203n9 1 3)MK176abn9 14)MK176,, 15)MKl O4,bn, 16)MK104,.

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Larnpiran 8. Profil pita DNA setiap primer

Keterangan : Pita DNA ke enam klon kelapa sawit hasil amplifikasi menggunakan primer (a) OPN-18; (b) OPN-16; (c) OPC-09; Lajur (M) 1 kb DNA Ladder, 1)MKl 52j, 2)MKl 52abn, 3)MK152,, 4)MK2Ogj, 5)MK2Ogabn, 6)MK209,, 7)MK2 1 2j, 8)MK2 12ah, 9)MK212,, 1 O)MK203j, 1 l)MK203,bn, 12)MK203,, 13)MKl 76,bn, 14)MK176,, 15)MKl O4,bn, 16)MK104,.

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Lanjutan

(d)OPW-19; (e) OPC-08;

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OPC-02; Lajur (M) 1 kb DNA Ladder, l)MK152j, 2)MKl 52ah, 3)MK152,, 4)MK2Ogj, 5)MK2O9abn, 6)MK209n, 7)MK21 2j, 8)MK212abn, 9)MK212,, 1 0)MK203j, 1 l)MK203abn, 12)MK203,, 13)MK176,h, 14)MK176,, 15)MKl O4,bn, 16)MK104,.

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Keterangan : Pita DNA 6 klon kelapa sawit hasil amplifikasi menggunakan primer

(g)OPC-07; (h)OPA-02; (i)OPB-06; Lajur (M) 1 kb DNA Ladder, l)MK152j, 2)MKl 52ah, 3)MK152,, 4)MK2Ogj, 5)MK209abn, 6)MK209,,

.

7)MK212j, 8)MK2 1 2,bn, 9)MK212,, 1 O)MK203j, 1 l)MK203abn, 12)MK203,, 13)MK176,h, 14)MK176,, 15)MKl 04,h, 16)MK104,.

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Cj)OPB-05; (k)SC 10-20; (1)OPA-04; Lajur (M) 1 kb DNA Ladder, 1)MKl 52j, 2)MK152,h, 3)MK152,, 4)MK2Ogj, 5)MK2Ogah, 6)MK209,, 7)MK2 1 2j, 8)MK212,h, 9)MK212,, 1 0)MK203j, 1 l)MK203,h, 12)MK203,, 13)MKl 76,bn, 14)MK176,, 15)MKl 04ab,, 16)MK104,.