AlanFung:Protocol/KAPA Library Quantification Kits-GA: Difference between revisions

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==qPCR Cycling==
Step1 95C 5M
Step2 95C 30S
Step3 60C 45S
Step4 Plate Read
Step5 Go to Step 2, Repeat 34X

Revision as of 18:58, 20 January 2010

Prepare qPCR Reagents

Ensure that the following reagents are completely thawed and thoroughly mixed by vortexing

  • 2X KAPA SYBR FAST qPCR Master Mix
  • 6 DNA Standards
  • 1:1000, 1:2000, 1:4000 & 1:8000 dilution of library DNA

Step1

  • Perform an an 1:1000 dilution of the purified library in Library Dilution Buffer
  • Optional 1:2000, 1:4000, 1:8000

Step2

qPCR Setup

Setup Volume(ul)
KAPA SYBR FAST qPCR Master Mix Containing Primer Premix 12
PCR-grade water 4
Diluted library DNA or DNA Standard (1-6) 4

qPCR Cycling

Step1 95C 5M
Step2 95C 30S
Step3 60C 45S
Step4 Plate Read
Step5 Go to Step 2, Repeat 34X