Richard Lab:mRNA quantification: Difference between revisions
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(New page: This page is for mRNA quantification; ===Reverse Transcription=== 1.Prepare the foloowing reaction mix (in this order): ::*2μL Nuclease free water ::*1μL Random hexamer mix (60μM) ::*5...) |
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Revision as of 11:56, 26 October 2011
This page is for mRNA quantification;
Reverse Transcription
1.Prepare the foloowing reaction mix (in this order):
- 2μL Nuclease free water
- 1μL Random hexamer mix (60μM)
- 5μL AMV reaction mix
- 1μL RNA from extraction (500-1000ng/μL)
- 1μL AMV reverse transcriptase
2.Incubate at 25°C for 5 min. 3.Incubate at 42°C for 1 hour. 4.Inactivate enzyme at 80°C for 5 minutes. 5.Add 20μL nuclease free water. 6.Store at -20°C.
