The following DNased RNA samples showed inconsistencies between qPCR reps (one rep showed amplification, the other rep did not) on 20150514:
NC1
SC1
SC2
SC4
Reran these four samples to obtain a definitive answer as to whether or not they have residual gDNA in them prior to using them to make cDNA.
Used Oly_Actin primers (SR IDs: 1504, 1505)
Used 1μL from all templates.
All samples were run in duplicate.
Positive control was HL1 O.lurida DNA isolated by Jake on 20150323.
Cycling params:
95C – 2.5mins
40 cycles of:
95C – 10s
60C – 20s
Melt curve
Master mix calcs: 20150521_qPCR_Oly_DNased_RNA
Plate layout: 20150521_qPCR_plate_Jake_Oly_DNased_RNA
Results:
qPCR Data File (Opticon): Sam_20150521_145749.tad qPCR Report (Google Sheet): 20150521_qPCR_Report_Jake_Oly_DNased_RNA
No amplification in any of the RNA samples, nor the NTCs. Will make cDNA.
Amplification Plots
(http://eagle.fish.washington.edu/Arabidopsis/20150521_qPCR_Amp_Jake_Oly_DNased%20RNA_.JPG)
Melt Curves
(http://eagle.fish.washington.edu/Arabidopsis/20150521_qPCR_Melt_Jake_Oly_DNased%20RNA_.JPG)