Inoculated 3 x 5mL Marine Broth + biphenyl crystals (UV sterilized) cultures from frozen stock in 50mL Falcon tubes. Incubated 200RPM, 28C. Will use these are starter cultures to inoculate larger cultures.
qPCR – MV hemocyte cDNA from 20090614
Set up qPCR with Cv_CIAPIN_F/R primers. This is a second rep. Plate layout/PCR set up is here.
Results: Waters are clean. To be analyzed later with other genes that Mac has run.
qPCR – MV hemocyte cDNA from 20090614
Set up qPCR with Cv_CatY_F/R primers. This is a second rep. Plate layout/PCR set up is here.
Results: Waters are clean. To be analyzed later with other genes that Mac has run.
qPCR – MV hemocyte cDNA from 20090614
Set up qPCR with Cv_CatL_F/R primers. This is a second rep. Plate layout/PCR set up is here.
Results: Waters are clean. To be analyzed later with other genes that Mac has run.
qPCR – MV hemocyte cDNA from 20090614
Set up qPCR with Cv_TLR_”short”_F/R primers. This is a second rep. Plate layout/PCR set up is here.
Results: Waters are clean. To be analyzed later with other genes that Mac has run.
qPCRs – MV hemocyte cDNA from 20090614
qPCR – BgBL Primers
Set up qPCR with Cv_BgBL_F/R primers. This is a second rep. Plate layout/PCR set up is here.
Results: Waters are clean. To be analyzed later with other genes that Mac has run.
qPCR – HMG Primers
Set up qPCR with Cv_HMG_F/R primers. This is a second rep. Plate layout/PCR set up is here.
Results: Waters are clean. To be analyzed later with other genes that Mac has run.
qPCR – HSP70 Primers
Set up qPCR with Cv_HSP70_F/R primers. This is a second rep. Plate layout/PCR set up is here.
Results: Waters are clean. To be analyzed later with other genes that Mac has run.
qPCRs – MV hemocyte cDNA from 20090614
qPCR – CysB Primers
Set up qPCR with Cv_CysB_F/R. Plate layout/PCR set up is here.
Results: Waters are clean. To be analyzed later with other genes that Mac has run.
qPCR -BgBp Primers
Set up qPCR with Cv_BgBp_F/R primers. Plate layout/PCR set up is here.
Results: Waters are clean. To be analyzed later with other genes that Mac has run.
qPCR – MV hemocyte cDNA from 20090614
Set up qPCR with Cv_18s_F/R primers. Plate layout/PCR set up is here. This is a second rep. Used 1:20 cDNA plate made by Mac as template.
Results: Waters are clean. Still some samples that aren’t coming up. Will eventually run those samples using undiluted cDNA. To be analyzed later with other genes that Mac has run.
qPCR – Abalone Dg DNased RNA from yesterday and earlier today
Check Abalone RNA for residual gDNA contamination after DNase treatment. Ran qPCR with H.crach_16s_SYB_F/R primers. Plate layout/PCR set up here.
Results: Still gDNA in virtually every sample! This is totally insane. Will find/design primers that will only amplify gDNA to aid in subsequent analysis…
DNase Treatment – Abalone Dg RNA
DNase treated Abalone Dg RNA. Followed Ambion’s Turbo DNA-free Kit rigorous protocol. Here are the calcs/workup for the DNase treatment. This used previously untreated RNA.