Richard Lab:mRNA quantification

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This page is for mRNA quantification;
This page is for mRNA quantification;
===RNA extraction===
===RNA extraction===
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1.Use a kit or take a lookt at [[RNA extraction|this hub page]]
+
1.Use a kit or take a lookt 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.
===You can use a a reverse transcription PCR for this  
===You can use a a reverse transcription PCR for this  

Revision as of 16:56, 29 October 2011

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.

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