Daniel:Notebook/PosSequencing/2017-5-24: Difference between revisions
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>Djacobse (Created page with "=Barcode Oligo Production= Back to Calendar Making more double stranded barcode oligo since I'm running out. I'll make both EcoRI and Uraci...") |
>Djacobse |
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| width=" | | width="210" height="30" | Reagent | ||
| width="85" | uL Single rxn | | width="85" | uL Single rxn | ||
| width="95" | Master Mix Eco1 (8.2X) | | width="95" | Master Mix Eco1 (8.2X) |
Revision as of 15:17, 25 May 2017
Barcode Oligo Production
Making more double stranded barcode oligo since I'm running out. I'll make both EcoRI and Uracil versions.
Protocol
- qPCR
- Make the following master mixes
- Add 50 uL (dual) to the appropriate lanes File:PlateLayout-20170520-SinglePrimer.png
- qPCR Cycles
- 95C 3 min
- 95C 3 sec
- 55C 30 sec
- 72C 20 sec
- plate read
- goto b x30
- 72C 2 min
- 16C hold
Reagent | uL Single rxn | Master Mix Eco1 (8.2X) | Master Mix Eco2 (8.2X) | Master Mix Eco3 (8.2X) | Master Mix BioU1 (8.2X) | Master Mix BioU2 (8.2X) | Master Mix BioU3 (8.2X) |
Biotin-Uracil Primer (10 uM) | 1 | 0 | 0 | 0 | 8.2 | 8.2 | 8.2 |
Biotin-EcoRI Primer (10 uM) | 1 | 8.2 | 8.2 | 8.2 | 0 | 0 | 0 |
Primer 2 (10 uM) | 1 | 8.2 | 8.2 | 8.2 | 8.2 | 8.2 | 8.2 |
Template Oligo (100 pM) | 2 | 16.4 | 16.4 | 16.4 | 16.4 | 16.4 | 16.4 |
Kapa 2X Master Mix | 25 | 205 | 205 | 205 | 205 | 205 | 205 |
nfH2O | 21 | 172.2 | 172.2 | 172.2 | 172.2 | 172.2 | 172.2 |
Total | 50 | 410 | 410 | 410 | 410 | 410 | 410 |