Daniel:Notebook/PosSequencing/2017-6-13: Difference between revisions
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>Djacobse (Created page with "=Biotin vs Dual Biotin= Back to Calendar '''Workflow Reminder''' 350x265px ==Prot...") |
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<li>Prepare 2 plates; 1 single biotin and 1 dual biotin</li> | <li>Prepare 2 plates; 1 single biotin and 1 dual biotin</li> | ||
<li>For each sample prepare 58X master mix and (4X) master mix: 1X components set to 30 uL; add 2uL sample</li> | <li>For each sample prepare 58X master mix and (4X) master mix: 1X components set to 30 uL; add 2uL sample</li> | ||
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<li>58 (4) uL 1 uM primer2</li> | <li>58 (4) uL 1 uM primer2</li> | ||
<li>(4) uL 1 uM biotin/2biotin uracil primer</li> | <li>(4) uL 1 uM biotin/2biotin uracil primer</li> |
Revision as of 00:02, 13 June 2017
Biotin vs Dual Biotin
Workflow Reminder
File:ExperimentMap-20170610-Biotin-DualBiotin.png
Protocol
- qPCR
- Prepare 2 plates; 1 single biotin and 1 dual biotin
- For each sample prepare 58X master mix and (4X) master mix: 1X components set to 30 uL; add 2uL sample
- 58 (4) uL 1 uM primer2
- (4) uL 1 uM biotin/2biotin uracil primer
- 58 uL 10 pM barcode oligo v2
- 870 (60) uL 2X Kapa SYBR
- 638 (44) uL nfH2O
- Add 28 uL master mix according to plate (4X master mix is for positive controls, 58X is for samples and negative controls)
- Add 2 uL appropriate sample according to plate layout
- Use "standard" thermocycling program