Daniel:Notebook/HiResChrPaint/2014-9-10
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Padlock Probes II
Nanodrop Results
DpnII/USER Digestion
Reaction Table
Reagent | PPRL (uL) |
amplicon | 30 |
ug Input | 2.0 |
USER | 5 |
nf H2O | 45 |
Total | 80 |
- Set up reaction according to table in a 0.2 mL PCR tube
- Incubate for 2 hours at 37C
- Add 15 uL according to DpnII Primer table (below)
DpnII Primer Table
Reagent | PPRL (uL) |
10X DpnII Buffer | 10 |
100 uM RE-DpnII-V4 guide oligo | 5 |
Total | 15 |
- Incubate at 94C for 2 minutes
- Cool to 37C and incubate for 3 minutes
- Add 5 uL DpnII and incubate at 37C for 2 hours
- Heat inactivate DpnII at 65C for 20 minutes
Nanodrop Results
Sample | ng/uL ssDNA | ug in 20 uL | Total |
PPRL-DpnII-A | 38.7 | 0.8 | |
PPRL-DpnII-B | 35.3 | 0.7 | 1.5 |