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

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(Created page with '==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:…')
 
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==Step2==
==Step2==
==qPCR Setup==
{| {{table}}
| 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
|-
|
|}

Revision as of 18:55, 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