Daniel:Notebook/PosSequencing/2017-6-13: Difference between revisions
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>Djacobse |
>Djacobse |
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<li>Use "standard" thermocycling program</li> | <li>Use "standard" thermocycling program</li> | ||
</ol> | </ol> | ||
==Results== | |||
<gallery perrow=2 heights=250px widths=350px mode=packed-hover caption="Single Biotin Results"> | |||
File:PlateLayout-20170613-BiotinTest.png|Plate array | |||
File:Singlebiotin-20170613-plateCTs.png|Plate CTs | |||
File:Singlebiotin-20170613-plateCTs-normed.png|Normalized plate CTs | |||
File:Singlebiotin-20170613-rawcurves.png|Raw curves | |||
</gallery> | |||
<gallery perrow=2 heights=250px widths=350px mode=packed-hover caption="Single Biotin Results"> | |||
File:PlateLayout-20170613-BiotinTest.png|Plate array | |||
File:Dualbiotin-20170613-plateCTs.png|Plate CTs | |||
File:Dualbiotin-20170613-plateCTs-normed.png|Normalized plate CTs | |||
File:Dualbiotin-20170613-rawcurves.png|Raw curves | |||
</gallery> |
Revision as of 17:01, 14 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
- PlateLayout-20170613-BiotinTest.png
Plate array
- Singlebiotin-20170613-plateCTs.png
Plate CTs
- Singlebiotin-20170613-plateCTs-normed.png
Normalized plate CTs
- Singlebiotin-20170613-rawcurves.png
Raw curves
- PlateLayout-20170613-BiotinTest.png
Plate array
- Dualbiotin-20170613-plateCTs.png
Plate CTs
- Dualbiotin-20170613-plateCTs-normed.png
Normalized plate CTs
- Dualbiotin-20170613-rawcurves.png
Raw curves