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First Strand cDNA Synthesis (AB Kit)

795 bytes added, 16:27, 1 June 2012
Wrote protocol for 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:

{| border="1"
|+ Master Mix Components
! scope="col" | Component
! scope="col" | Per Reaction
! scope="col" | Per 10 Reactions
|-
! scope="row" | 10X RT Buffer
| 2 || 20
|-
! scope="row" | 25X dNTP Mix (100 mM)
| 0.8 || 8
|-
! scope="row" | 10X RT Random Primers
| 2 || 20
|-
! scope="row" | MultiScribe Reverse Transcriptase
| 1 || 10
|-
! scope="row" | 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'''. Takes ~2h