Ran qPCR on DNased RNA from earlier today to assess whether there was any residual gDNA after the DNase treatment with Oly_Actin_F/R primers (SR IDs: 1505, 1504).
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 are here: 20150514_qPCR_Oly_DNased_RNA
qPCR Plate Layout: 20150514_qPCR_plate_Jake_Oly_Control_RNA
Results:
qPCR Data File (Opticon): Sam_20150514_153529.tad
qPCR Report (Google Spreadsheet): 20150514_qPCR_Report_Jake_Oly_DNased_Control_RNA
Positive control comes up around cycle ~21.
No amplification in the no template controls.
Four wells of the DNased RNA samples exhibit amplification (B5, C10, C12, D3), however each respective replicate does not. Will re-test these four samples (NC1, SC1, SC2, SC4).
Amplification Plots
(http://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Amp_DNased_RNA_Jake_Oly_controls.JPG)
Melt Curves
(http://eagle.fish.washington.edu/Arabidopsis/20150514_qPCR_Melt_DNased_RNA_Jake_Oly_controls.JPG)