{"id":2790,"date":"2017-08-10T14:17:47","date_gmt":"2017-08-10T14:17:47","guid":{"rendered":"https:\/\/doktermuslim.com\/?p=2790"},"modified":"2017-08-10T14:17:47","modified_gmt":"2017-08-10T14:17:47","slug":"protokol-isolasi-rna-dengan-kit-geneaid","status":"publish","type":"post","link":"https:\/\/doktermuslim.com\/protokol-isolasi-rna-dengan-kit-geneaid\/","title":{"rendered":"Protokol Isolasi RNA dengan Kit Geneaid"},"content":{"rendered":"

Protokol Isolasi RNA dengan Kit geneaid sering digunakan dalam penelitian modern bioteknologi. Isolasi RNA merupakan langkah awal dalam memperoleh RNA dari jaringan, yang kemudian dapat kita periksa dengan qPCR baik realtime qPCR maupun PCR konvensional. Reagen yang dipakai : Kit RNA Mini Tissue Geneaid<\/a><\/p>\n

Persiapan Protokol Isolasi RNA dengan Kit Geneaid<\/u><\/strong><\/h3>\n

A. Persiapan Kit<\/em><\/strong><\/h4>\n
    \n
  1. Tambahkan 100 ml ethanol absolut<\/li>\n
  2. masukkan ke dalam ml wash buffer, campur sampai homogen.<\/li>\n
  3. Beri tanda V dan tanggal penambahan ethanol absolut<\/li>\n
  4. ambil 2500 mikro liter RB buffer, masukkan dalam konikal 15 ml<\/li>\n
  5. tambahkan 25 mikro liter Beta Mercapto ethanol<\/li>\n
  6. Campur, sampai homogen, beri label, tutup dengan aluminium foil<\/li>\n
  7. ambil 700 mikro liter\/7 ml ethanol100%, lalu masukkan ke dalam konikal 15 ml<\/li>\n
  8. tambahkan 3 ml RNAse, DNAse free water<\/li>\n
  9. campur hingga homogen dan berilah label.<\/li>\n<\/ol>\n

    B. Persiapan Sampel<\/em><\/strong><\/h4>\n
      \n
    1. Keluarkan sampel jaringan dari kulkas suhu -80 derajat celcius.<\/li>\n
    2. potong jaringan dengan lancet<\/li>\n
    3. timbang jaringan sekitar 25 mg, lalu masukkan ke dalam tube 1,5 ml<\/li>\n<\/ol>\n

      Protokol Isolasi RNA<\/u><\/strong><\/h3>\n
        \n
      1. Jaringan 25 mg, tambahkan 400 mikro liter RB buffer yang sudah ditambah dengan Beta Mercapto ethanol<\/li>\n
      2. lakukan homogenisasi dengan micropastle sampai benar-benar homogen.<\/li>\n
      3. Pindahkan ke filter collum dan injubasi selama 3 menit<\/li>\n
      4. sentrifugasi 1000 g, selama 30 detik.<\/li>\n
      5. pindahkan filtrat ke dalam tube 1,5 ml dan tambahkan ethanol 70% sebanyak 400 mikro liter.<\/li>\n
      6. campur dengan di gosok<\/li>\n
      7. transfer larutan ke dalam RB collum dan dilanjutkan sentrifugasi 16000 g selama 1 menit.<\/li>\n
      8. Buang supernatan, lalu bagian tube atas yang terdapat filter, tambahkan 400 mikro liter W1 buffer ke dalam RB collum<\/li>\n
      9. sentrifugasi 16.000 g selama 30 detik, dan buang supernatan.<\/li>\n
      10. tambahkan 600 mikro liter wash buffer ke dalam RB column dan sentrifugasi 16.000 g selama 30 detik.<\/li>\n
      11. buang supernatan, dan tambahkan 600 mikroliter wash buffer lagi ke dalam RB column<\/li>\n
      12. sentrifugasi dengan kecepatan 16000 g selama 30 detik, dan buang sipernatan.<\/li>\n
      13. sentrifugasi lagi 16.000 g selama 3 menit tanpa menambah apapun.<\/li>\n
      14. gantilah collection tube dengan tube 1,5 ml<\/li>\n
      15. tambahkan RNAse free water sebanyak 50 mikro liter.<\/li>\n
      16. inkubasi selama 10 menit dengan suhu ruangan<\/li>\n
      17. sentrifugasi 16.000 g selama 1 menit<\/li>\n
      18. pindahkan eluen ke dalam tube 1,5 ml yang baru<\/li>\n<\/ol>\n","protected":false},"excerpt":{"rendered":"

        Protokol Isolasi RNA dengan Kit geneaid sering digunakan dalam penelitian modern bioteknologi. Isolasi RNA merupakan langkah awal dalam memperoleh RNA dari jaringan, yang kemudian dapat [Read more]<\/a><\/p>\n","protected":false},"author":1,"featured_media":2791,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[13],"tags":[1671,1670,1674,1672,1673],"newstopic":[],"yoast_head":"\nProtokol Isolasi RNA dengan Kit Geneaid | Dokter Muslim<\/title>\n<meta name=\"description\" content=\"Protokol Isolasi RNA dengan Kit geneaid sering digunakan dalam penelitian modern bioteknologi. Reagen yang dipakai : Kit RNA Mini Tissue Geneaid.\" \/>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/doktermuslim.com\/protokol-isolasi-rna-dengan-kit-geneaid\/\" \/>\n<meta property=\"og:locale\" content=\"id_ID\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Protokol Isolasi RNA dengan Kit Geneaid | Dokter Muslim\" \/>\n<meta property=\"og:description\" content=\"Protokol Isolasi RNA dengan Kit geneaid sering digunakan dalam penelitian modern bioteknologi. 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