BAB VI KESIMPULAN DAN SARAN
6.2 Saran
1. Perlu adanya kombinasi metode guna verifikasi lebih lanjut mengenai analisis cemaran daging babi dengan menggunakan metode PCR, misalnya dengan menggunakan sequencing dan Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP).
2. Perlu dilakukan inovasi pengembangan metode PCR dalam analisis DNA babi pada sampel makanan, seperti pembuatan kit spesifik untuk mendeteksi daging babi, sehingga proses analisis dapat lebih cepat.
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Lampiran 1
Tabel 4. Konsentrasi dan kemurnian hasil isolasi genom
No Sampel Konsentrasi (ng/μl) Kemurnian (A260/A280) 1 Daging babi 960,8 1,52 2 Daging sapi 1280,5 1,07 3 0,1% daging babi 1655,1 1,71 4 0,5% daging babi 1845,3 1,73 5 1% daging babi 839,1 1,62 6 2,5% daging babi 901,7 1,65 7 5% daging babi 1465,6 1,74 8 10% daging babi 1640,1 1,76
9 Daging segar tanpa perlakuan 4797,3 1,08 10 Daging segar dengan pemanasan 4826,6 1,05 11 Daging segar dicuci dengan n-hekssan 4720,5 1,33 12 Daging segar dicuci dengan air 4687,3 1,45 13 Sampel tanpa perlakuan 1588,3 1,91 14 Sampel dengan pemanasan 1752,0 1,87 15 Sampel dicuci dengan n-heksan 2812,7 1,84 16 Sampel dicuci dengan air 1966,4 1,85
17 Sampel 1 1615,8 1,87 18 Sampel 2 1308,6 1,60 19 Sampel 3 2790,3 1,70 20 Sampel 4 1018,7 1,66 21 Sampel 5 1007,2 1,69 22 Sampel 6 1794,8 1,88
Lampiran 2
Kondisi PCR yang digunakan untuk amplifikasi menggunakan primer spesifik DNA sapi dan primer spesifik DNA babi adalah sama, yaitu:
Gambar 15. Kondisi PCR
Lampiran 3
Tabel 5. Komposisi campuran reaksi PCR
Komposisi Jumlah Buffer PCR 10x 5 µl MgCl Buffer 3mM 5 µl dNTP 200µM 4 µl Taq Polymerase 0,4 µl Primer Forward 10µM 2 µl Primer Reverse 10µM 2 µl Template 2 µl ddH2O 29,6 µl Total 50 µl 940 C 610 C 940 C 720 C 720 C 40 C 5 menit ~ 5 menit 90 detik 45 detik 45 detik 35 siklus
Lampiran 4.
Tabel 6. Komposisi bahan yang digunakan
Larutan Komposisi dan cara pembuatan Sumber EDTA (0,5 M; pH 8,0) Sebanyak 186,1 g disodium
EDTA.2H2O dilarutkan dalam 800 ml H2
Sambrook & Russell, 2001: A1.26
O, dihomogenkan dengan menggunakan magnetic stirrer,
larutan ditambahkan NaOH hingga pH 8,0
Tris-Cl (10 mM; pH 8,0) 121,1 g tris dilarutkan ke dalam 800 ml H2 Sambrook & Russell, 2001: 6.28 0, pH disesuaikan menggunakan HCl hingga pH 8,0 TE buffer (pH 8,0) 100 mM Tris pH 8,0 10 mM EDTA pH 8,0 Sambrook & Russell, 2001: A1.7
TAE 50X Sebanyak 242 g tris; 57,1ml asam asetat glasial; dan 100 ml EDTA 0,5M (pH 8,0) dilarutkan dalam air hingga volume 1 L
Sambrook & Russell, 2001: A1.17
Cell Buffer Lysis 10 mM Tris-Cl (pH 8,0)
1m M EDTA (pH8,0) 1% (w/v) SDS Sambrook & Russel, 2001: 6.28; Kesmen, 2009 NaCl 5 M 100 ml 29,29g NaCl dilarutkan dalam 100 ml
H2O Kloroform-isoamil
alkohol (24:1), 100 ml
96 ml kloroform 4 ml isoamil alkohol
