Revision as of 00:50, 11 September 2014 by >Djacobse
Padlock Probes II
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Nanodrop Results
Sample
|
1:1 ng/uL
|
5:1 ng/uL
|
5:1 ng/uL Projected
|
10:1 ng/uL
|
10:1 ng/uL Projected
|
Average ng/uL
|
Standard Deviation
|
ug in 100 uL
|
PPRL-A
|
1944.1
|
373
|
1865
|
184.4
|
1844
|
1884.4
|
52.8
|
188.4
|
PPRL-B
|
2064.8
|
421.7
|
2108.5
|
203
|
2030
|
2067.8
|
39.3
|
206.8
|
Usually after ethanol precipitation I get really high results, so I'll just say these are fine and continue as though I have a reasonable amount of material.
DpnII/USER Digestion
Reaction Table
Reagent
|
PPRL-A1 (uL)
|
PPRL-A2 (uL)
|
PPRL-B1 (uL)
|
PPRL-B2 (uL)
|
amplicon
|
50
|
50
|
50
|
50
|
ug Input
|
94.2
|
94.2
|
103.4
|
103.4
|
USER
|
5
|
5
|
5
|
5
|
nf H2O
|
30
|
30
|
30
|
30
|
Total
|
85
|
85
|
85
|
85
|
- Set up reaction according to table in a 0.2 mL PCR tube
- Incubate for 2 hours at 37C
- Add 10 uL DpnII buffer (10X)
- 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
- [[Daniel:Protocols/
Nanodrop Results
Sample
|
PPRL-A1
|
PPRL-A2
|
PPRL-B1
|
PPRL-B2
|
ng/uL
|
154.1
|
131
|
153
|
163.7
|
ug in 40 uL
|
6.2
|
5.2
|
6.1
|
6.5
|
ug total
|
|
|
|
24.1
|
TBE Gel
Lane
|
1
|
2
|
3
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4
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5
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6
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7
|
8
|
9
|
10
|
11
|
12
|
Sample
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