Reverse Transcription – Herring Liver mRNA for SOLiD Libraries

Ligation reactions from yesterday were subject to reverse transcription according to protocol. Master mix workup info is here. Samples were incubated @ 42C, 30mins and then cleaned up using the Qiagen MiniElute PCR Purification Kit, according to the Ambion WTK protocol.

After RT rxn, samples were run on a Novex 6% TBE-Urea gel according to Ambion WTK protocol. Samples were loaded, left to right: 2L, 3L, 4L, 6L. Ladders are to the left of each sample. The break in the smear in the 6L sample is a tear in the gel.

The recommended range of cDNA (100-200bp) were excised from the gel and were cut into four pieces, according to the Ambion WTK protocol. The two outer gel slices from each sample will be stored @-20C. Then proceeded to the emulsion PCR. Here is an image of the gel after the cDNA was cut out:

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