Daniel:Notebook/HiResChrPaint/2013-6-5: Difference between revisions
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2. Incubate in thermocycler at 37C for 16 hours; heat shock 20 minutes at 65 C | 2. Incubate in thermocycler at 37C for 16 hours; heat shock 20 minutes at 65 C | ||
3. [[Daniel:Protocols/Qiaquick_Column|Column purification]], elute with 30 uL EB | |||
4. Measure in Nanodrop | |||
===Nanodrop Results=== | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt;font-weight:bold" valign="bottom" | |||
| width="65" height="15" | Sample | |||
| width="65" | a-4C | |||
| width="65" | a-6C | |||
| width="65" | b-4C | |||
| width="65" | b-6C | |||
| width="65" | a-4D | |||
| width="65" | a-6D | |||
| width="65" | b-4D | |||
| width="65" | b-6D | |||
|- style="font-size:12pt" valign="bottom" | |||
|style="font-weight:bold" height="15" | ng/uL | |||
| align="center" | 119.1 | |||
| align="center" | 119.2 | |||
| align="center" | 108.5 | |||
| align="center" | 140.4 | |||
| align="center" | 121.8 | |||
| align="center" | 147.7 | |||
| align="center" | 112.2 | |||
| align="center" | 128.7 | |||
|- style="font-size:12pt" valign="bottom" | |||
|style="font-weight:bold" height="15" | ug in 30 uL | |||
| align="center" | 3.6 | |||
| align="center" | 3.6 | |||
| align="center" | 3.3 | |||
| align="center" | 4.2 | |||
| align="center" | 3.7 | |||
| align="center" | 4.4 | |||
| align="center" | 3.4 | |||
| align="center" | 3.9 | |||
|- style="font-size:12pt" valign="bottom" | |||
|style="font-weight:bold" height="15" | Yield (%) | |||
| align="center" | 78.0 | |||
| align="center" | 73.7 | |||
| align="center" | 80.1 | |||
| align="center" | 85.2 | |||
| align="center" | 80.1 | |||
| align="center" | 81.6 | |||
| align="center" | 77.0 | |||
| align="center" | 77.1 | |||
|} | |||
===TBE Gel=== | |||
Check to make sure samples are at least mostly digested. |
Revision as of 22:48, 5 June 2013
Probe Prep 3 (Started 5/29/2013)
DpnII Digestion
Finishing the digest on the PCR amplified probes, aa-dUTP and no aa-dUTP. Will run for 2 hours since I didn't see any qualitative difference between the two.
1. Set up
C group
Sample | uL sample | uL Dpn II (50 U/uL) | uL 10x buffer | uL H20 | total |
alpha 4-C | 35 | 2 | 5 | 8 | 50 |
alpha 6-C | 35 | 2 | 5 | 8 | 50 |
beta 4-C | 35 | 2 | 5 | 8 | 50 |
beta 6-C | 35 | 2 | 5 | 8 | 50 |
D Group
Sample | uL sample | uL Dpn II (50 U/uL) | uL 10x buffer | uL H20 | total |
alpha 4-D | 35 | 2 | 5 | 8 | 50 |
alpha 6-D | 35 | 2 | 5 | 8 | 50 |
beta 4-D | 35 | 2 | 5 | 8 | 50 |
beta 6-D | 35 | 2 | 5 | 8 | 50 |
2. Incubate in thermocycler at 37C for 16 hours; heat shock 20 minutes at 65 C 3. Column purification, elute with 30 uL EB 4. Measure in Nanodrop
Nanodrop Results
Sample | a-4C | a-6C | b-4C | b-6C | a-4D | a-6D | b-4D | b-6D |
ng/uL | 119.1 | 119.2 | 108.5 | 140.4 | 121.8 | 147.7 | 112.2 | 128.7 |
ug in 30 uL | 3.6 | 3.6 | 3.3 | 4.2 | 3.7 | 4.4 | 3.4 | 3.9 |
Yield (%) | 78.0 | 73.7 | 80.1 | 85.2 | 80.1 | 81.6 | 77.0 | 77.1 |
TBE Gel
Check to make sure samples are at least mostly digested.