Daniel:Notebook/ComboLock/2016-11-16: Difference between revisions
Jump to navigation
Jump to search
>Djacobse |
>Djacobse |
||
Line 74: | Line 74: | ||
|- style="background-color:#C4BD97;font-size:12pt;font-weight:bold" align="center" | |- style="background-color:#C4BD97;font-size:12pt;font-weight:bold" align="center" | ||
| width="160" height="34" | Sample | | width="160" height="34" | Sample | ||
| width=" | | width="85" | ng/uL | ||
| width=" | | width="85" | ug in 30 uL | ||
|- style="font-size:12pt" | |- style="font-size:12pt" |
Revision as of 18:15, 16 November 2016
BSA Beads Test (Started Yesterday)
Protocol
- Rolling Circle Amplification
- Prepare 10.2X Master mix according to table below
- Add sample to new reaction tubes according to following table
- Incubate at 37C for 3 hours
- Incubate at 65C for 10 minutes;Hold at 10C until next step
- qPCR
Reagent | uL Added | Master Mix (10.2X) |
Template | 5 | NA |
RCA Primer (100 uM) | 2.5 | 25.5 |
dNTP (1 mM) | 0.8 | 8.16 |
10X Buffer | 2 | 20.4 |
Phi29 | 1 | 10.2 |
nfH2O | 8.7 | 88.74 |
Total | 20 | 153 |
Library Prep (Started Yesterday; Exp. Start 11/7/16)
- Ethanol Precipitation
- Centrifuge for 25 minutes at 12000 rpm at 4C
- Remove supernatant and add 750 uL 70% EtOH
- Centrifuge for 12 minutes at 12000 rpm at 4C
- Remove supernatant and spin in vacuum centrifuge for 10 minutes
- Add 30 (20 for NTC) uL nfH2O to resuspend; pool samples A and B by reusing the water
Sample | ng/uL | ug in 30 uL |
Cells RCA Product (2AB,2BB) |