Richard Lab:mRNA quantification
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This page is for mRNA quantification;
RNA extraction
1.Use a kit or take a lookt at this hub page</br> 2.Be really carefull when you do this, use DEPC water and some kind of RNAase degrading spray. ===You can use a a reverse transcription PCR for this
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.
