qPCR – RLO Prophage Genes

Ran qPCRs on a set of black abalone digestive gland DNA (sample list provided by Carolyn):

07:12-01 (Black Ab exp 1)
07:12-02 (Black Ab exp 1)
08:13-05 (Black Ab exp 2)
08:13-18 (Black Ab exp 2)
08:13-24 (Black Ab exp 2)
08:13-25 (Black Ab exp 2)
UW06:06-32
UW06:06-41

UW06:06-50 (Black Ab exp 1)
UW06:06-52 (Black Ab exp 1)

The two samples with a strikethrough did not have any DNA left in the tubes and were not run.

Gene targets:
– RLO Major Capsid Protein (RLO_MCP)
– RLO Prohead Protease Protein (RLO_ph_protease)
– XenoCal Phage Portal Gene (XC prophage)

Master mix calcs are here (Google Sheet): 20170413 – qPCR_XCphagePortal_RLOcapsid_RLOprohead

The same master mix calculations were used for each, just swapped in appropriate primers.

All samples were run in duplicate.

Cycling params, plate layout, etc. can be found in the qPCR Report (see Results below).

Results:
qPCR Report (PDF): Sam_2017-04-13 14-56-03_CC009827.pdf
qPCR Data File (CFX): Sam_2017-04-13 14-56-03_CC009827.pcrd

Melt curves for all three primer sets looked perfect (see below)

Amplification present for all samples, with all three primers except the 07:12 samples.

Will pass info along to Carolyn and Stan.

Will add info to the following two spreadsheets (Google Sheets):

Black Abalone: Expt 1 – WS & Phage

Black Abalone: Expt 2 – WS only

 


 

Green = RLO_ph_protease

Brown = RLO_MCP

Blue = XC_prophage

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