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)