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

from Sam’s Notebook http://ift.tt/1AnYtJp

qPCR – Re-run Jake’s O.lurida DNased RNA Samples NC1, SC1, SC2, SC4 from 20150514
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