Richard Lab:mRNA quantification: Difference between revisions
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This page is for mRNA quantification; | This page is for mRNA quantification; | ||
===RNA extraction=== | ===RNA extraction=== | ||
1.Use a kit or take a | 1.Use a kit or take a look at [[RNA extraction|this hub page]]<br> | ||
2.Be really carefull when you do this, use DEPC water and some kind of RNAase degrading spray. | 2.Be really carefull when you do this, use DEPC water and some kind of RNAase degrading spray.<br> | ||
You can use a a reverse transcription PCR for this.<br> | |||
===Reverse Transcription=== | ===Reverse Transcription=== | ||
Revision as of 13:57, 29 October 2011
This page is for mRNA quantification;
RNA extraction
1.Use a kit or take a look at this hub page
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.
