Tag Archives: SYTO13

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.

Share

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.

Share

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.

Share

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…

Share

qPCRs – MV hemocyte cDNA from 20090614

qPCR – CIAPIN Primers

Set up qPCR with Cv_CIAPIN_F/R primers. Plate layout/PCR set up is here.

Results: Waters are clean. gDNA samples did not amplify. To be analyzed later with other genes that Mac has run.

 

qPCR – CatY Primers

Set up qPCR with Cv_CatY_F/R primers. Plate layout/PCR set up is here.

Results: Waters are clean. gDNA samples did not amplify. To be analyzed later with other genes that Mac has run.

Share

qPCR – MV hemocyte cDNA: Test Immomix (SYTO13) vs. Strategene SYBR

Due to craziness seen in melting curves, fluorescence, and empty wells from the previous run, will compare SYTO vs. SYBR with select MV cDNAs. Additionally, acquired some qPCR strip caps to use instead of the ABI film. Used Cv_18s_F/R primers. qPCR set up/plate layout is here.

Results: Both seem to work fine. H2O fluorescence is weird, but doesn’t come up in the melting curves. Strategene SYBR provides a brighter signal, but results in a higher melting temp than the SYTO.

Share