First Strand cDNA Synthesis (AB Kit)

Materials

• Purified RNA
• High Capacity cDNA Reverse Transcription Kit without RNAse inhibitor (AB 4368813 for 1000 reactions, see http://products.invitrogen.com/ivgn/product/4368813)

Protocol

• Generate Master Mix Containing:

Master Mix Components

Component	Per Reaction	Per 10 Reactions
10X RT Buffer	2	20
25X dNTP Mix (100 mM)	0.8	8
10X RT Random Primers	2	20
MultiScribe Reverse Transcriptase	1	10
Nuclease Free Water	4.2	42

• Add 10 uL Master mix per well into tube or plate
• Add 0.1-10 ug RNA in a final volume of 10 uL to each well
• Run the PCR Program AB Reaction Protocol. Takes ~2h:
  • 10 min at 25C
  • 120 min at 37C
  • 5 min at 85C