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Isolation of RNA from blood

Posted by on April 30, 2014

Isolation of RNA from blood


 pdf icon2Isolation of RNA


  1. 500 µL of fresh blood put on eppendof 1.5 mL
  2. Added 500 µL tri  reagent, was pipetting
  3. Incubated for 5 minutes in romm temperature
  4. Added 100 µL chloroform
  5. shake vigorously for 15 seconds
  6. incubated  for 10-15 minutes at room temperature
  7. Centrifuged at 12,000 rpm 4 ° C for 15 minutes
  8. Taken / transfer aqueous phase ( in top layer ) to another eppendof 1.5 mL
  9. Added 250 µL isopropanol
  10. Incubated for 5-10 minutes at room temperature
  11. Centrifuged 12000 rpm 4 ° C for 8 minutes , the supernatant was discarded , was obtained the white pellet
  12. Added the Pellets with 500 µL 75 % ethanol
  13. Centrifuged 7500 rpm 4 ° C for 5 minutes, the supernatant was discarded completely , was obtained RNA pellet
  14. Briefly air-dry  the RNA pellets for 3-5 minutes remain in the laminar
  15. Added 20 µL of water free RNA
  16. The results are stored at -80 ° C for RT RNA or PCR process , in part to determine the levels and purity using the NanoDrop spectrometer

Note :
a. All procedures must doing in the laminary Air flow
b . All tools (eppendof , yellow type , blue type , white type of must have been in treatment with DEPC water treatment

One Response to Isolation of RNA from blood

  1. Habibi Ainun

    balik lagi ke lab :)

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