Daniel:Notebook/PosSequencing/2017-6-13: Difference between revisions
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>Djacobse |
>Djacobse |
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<li>870 (60) uL 2X Kapa SYBR</li> | <li>870 (60) uL 2X Kapa SYBR</li> | ||
<li>638 (44) uL nfH<sub>2</sub>O</li></ol> | <li>638 (44) uL nfH<sub>2</sub>O</li></ol> | ||
<li>Add 28 uL master mix according to plate (4X master mix is for positive controls, 58X is for samples and negative controls)</li> | <li>Add 28 uL master mix according to plate layout (4X master mix is for positive controls, 58X is for samples and negative controls)</li> | ||
[[Image:PlateLayout-20170613-BiotinTest.png|350x250px]] | |||
<li>Add 2 uL appropriate sample according to plate layout</li> | <li>Add 2 uL appropriate sample according to plate layout</li> | ||
<li>Use "standard" thermocycling program</li> | <li>Use "standard" thermocycling program</li> | ||
</ol> | </ol> |
Revision as of 18:48, 13 June 2017
Biotin vs Dual Biotin (Started Yesterday)
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 layout (4X master mix is for positive controls, 58X is for samples and negative controls) File:PlateLayout-20170613-BiotinTest.png
- Add 2 uL appropriate sample according to plate layout
- Use "standard" thermocycling program