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

Melt Curves