Sam’s Notebook: Samples Received – C.bairdi Hemolymph and Tissue in Ethanol from Pam Jensen

Pam Jensen (NOAA) brought by a variety of hemolymph and tissues stored in ethanol to use for DNA sequencing with the new MinION (Oxford Nanopore) to help aid our transcriptomics work.

The box of samples was stored in FTR 26 (underneath the fume hood).

Pam will provide a digitized version of the sample spreadsheet, as well as column name explanations. I’ll add that info here when I receive it.

Image of sample storage box

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Sam’s Notebook: qPCR – Geoduck hemolymph and hemocyte cDNA with vitellogenin primers

Previously isolated RNA on 20191125 and made cDNA on 20191126 from some geoduck hemolymph and hemocyte samples that Shelly asked me to run qPCRs on.

Ran qPCR with vitellogenin primers I previously designed on 20181129 and tested on 20181206

Primers:

SR IDs:

  • 1712 (Pg_DN51983i8_1471)
  • 1711 (Pg_DN51983i8_1347)

Positive control used was same pooled gonad cDNA used succesffully on 20181206 to test the assay. Due to sample limitations, positive control was not run in duplicate!

All qPCR reactions were run in duplicate. See qPCR Report (Results section below) for plate layout, cycling params, etc.

qPCR Master Mix calcs (Google Sheet):

NOTE: I actually ended up running this twice (55oC and 60oC)due to poor melt curves when run at 55oC anneal.