Daniel:Notebook/HiResChrPaint/2013-4-25: Difference between revisions
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[[Daniel:Notebook/HiResChrPaint|Back to calendar]] | [[Daniel:Notebook/HiResChrPaint|Back to calendar]] | ||
==DpnII Digest (Continued from [[Daniel:Notebook/HiResChrPaint/2013-4-24|4/24/2013]])== | |||
1. Took samples from thermocycler | |||
2. Purified using [[Daniel:Protocols/Qiaquick_Column|Qiagen column]], elute with 30 uL | |||
3. Set up following TBE gel | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt" align="center" | |||
| width="65" height="36" | Sample | |||
|style="background-color:#D9D9D9" width="65" | Ladder | |||
|style="background-color:#B7DEE8" width="65" | V4S3A-Pre Digestion | |||
|style="background-color:#B7DEE8" width="65" | V4S3B-Pre Digestion | |||
|style="background-color:#FABF8F" width="65" | V4S3A-Post Digestion | |||
|style="background-color:#CCC0DA" width="65" | V4S3A-Post Column | |||
|style="background-color:#CCC0DA" width="65" | V4S3B-Post Column | |||
|style="background-color:#D9D9D9" width="65" | Ladder | |||
|style="background-color:#B7DEE8" width="65" | V6S3A-Pre Digestion | |||
|style="background-color:#B7DEE8" width="65" | V6S3B-Pre Digestion | |||
|style="background-color:#FABF8F" width="65" | V6S3A-Post Digestion | |||
|style="background-color:#CCC0DA" width="65" | V6S3A-Post Column | |||
|style="background-color:#CCC0DA" width="65" | V6S3B-Post Column | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | uL sample | |||
|style="background-color:#D9D9D9" align="center" | 1 | |||
|style="background-color:#B7DEE8" align="center" | 2 | |||
|style="background-color:#B7DEE8" align="center" | 2 | |||
|style="background-color:#FABF8F" align="center" | 2 | |||
|style="background-color:#CCC0DA" align="center" | 2 | |||
|style="background-color:#CCC0DA" align="center" | 2 | |||
|style="background-color:#D9D9D9" align="center" | 1 | |||
|style="background-color:#B7DEE8" align="center" | 2 | |||
|style="background-color:#B7DEE8" align="center" | 2 | |||
|style="background-color:#FABF8F" align="center" | 2 | |||
|style="background-color:#CCC0DA" align="center" | 2 | |||
|style="background-color:#CCC0DA" align="center" | 2 | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | uL buffer | |||
|style="background-color:#D9D9D9" align="center" | 5 | |||
|style="background-color:#B7DEE8" align="center" | 4 | |||
|style="background-color:#B7DEE8" align="center" | 4 | |||
|style="background-color:#FABF8F" align="center" | 4 | |||
|style="background-color:#CCC0DA" align="center" | 4 | |||
|style="background-color:#CCC0DA" align="center" | 4 | |||
|style="background-color:#D9D9D9" align="center" | 5 | |||
|style="background-color:#B7DEE8" align="center" | 4 | |||
|style="background-color:#B7DEE8" align="center" | 4 | |||
|style="background-color:#FABF8F" align="center" | 4 | |||
|style="background-color:#CCC0DA" align="center" | 4 | |||
|style="background-color:#CCC0DA" align="center" | 4 | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | uL dye | |||
|style="background-color:#D9D9D9" align="center" | 2 | |||
|style="background-color:#B7DEE8" align="center" | 2 | |||
|style="background-color:#B7DEE8" align="center" | 2 | |||
|style="background-color:#FABF8F" align="center" | 2 | |||
|style="background-color:#CCC0DA" align="center" | 2 | |||
|style="background-color:#CCC0DA" align="center" | 2 | |||
|style="background-color:#D9D9D9" align="center" | 2 | |||
|style="background-color:#B7DEE8" align="center" | 2 | |||
|style="background-color:#B7DEE8" align="center" | 2 | |||
|style="background-color:#FABF8F" align="center" | 2 | |||
|style="background-color:#CCC0DA" align="center" | 2 | |||
|style="background-color:#CCC0DA" align="center" | 2 | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | loaded | |||
|style="background-color:#D9D9D9" align="center" | 7 | |||
|style="background-color:#B7DEE8" align="center" | 7 | |||
|style="background-color:#B7DEE8" align="center" | 7 | |||
|style="background-color:#FABF8F" align="center" | 7 | |||
|style="background-color:#CCC0DA" align="center" | 7 | |||
|style="background-color:#CCC0DA" align="center" | 7 | |||
|style="background-color:#D9D9D9" align="center" | 7 | |||
|style="background-color:#B7DEE8" align="center" | 7 | |||
|style="background-color:#B7DEE8" align="center" | 7 | |||
|style="background-color:#FABF8F" align="center" | 7 | |||
|style="background-color:#CCC0DA" align="center" | 7 | |||
|style="background-color:#CCC0DA" align="center" | 7 | |||
|} | |||
4. Ran gel for 30 minutes at 200 V. | |||
5. Stained for 10 minutes with 3 uL SYBR gold | |||
6. Measured concentration in Nanodrop | |||
7. Stored at 4C for exonuclease digestion tomorrow | |||
===Nanodrop Results=== | |||
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> | |||
|- style="font-size:12pt" align="center" | |||
| width="65" height="15" | Sample | |||
| width="65" | ng/uL | |||
| width="65" | uL | |||
| width="65" | ng | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | V4S3A | |||
| align="center" | 102.2 | |||
| align="center" | 30 | |||
| align="center" | 3066 | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | V4S3B | |||
| align="center" | 86.8 | |||
| align="center" | 30 | |||
| align="center" | 2604 | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | V6S3A | |||
| align="center" | 80.1 | |||
| align="center" | 30 | |||
| align="center" | 2403 | |||
|- style="font-size:12pt" align="center" | |||
| height="15" | V6S3B | |||
| align="center" | 77.9 | |||
| align="center" | 30 | |||
| align="center" | 2337 | |||
|} | |||
===Gel Results=== | |||
[[Image:DpnII_Digest_042513.jpg|600px]] | [[Image:DpnII_Digest_042513.jpg|600px]] |
Latest revision as of 19:01, 26 April 2013
Probe Prep Test (Started 4/22/2013)[edit]
DpnII Digest (Continued from 4/24/2013)[edit]
1. Took samples from thermocycler 2. Purified using Qiagen column, elute with 30 uL 3. Set up following TBE gel
Sample | Ladder | V4S3A-Pre Digestion | V4S3B-Pre Digestion | V4S3A-Post Digestion | V4S3A-Post Column | V4S3B-Post Column | Ladder | V6S3A-Pre Digestion | V6S3B-Pre Digestion | V6S3A-Post Digestion | V6S3A-Post Column | V6S3B-Post Column |
uL sample | 1 | 2 | 2 | 2 | 2 | 2 | 1 | 2 | 2 | 2 | 2 | 2 |
uL buffer | 5 | 4 | 4 | 4 | 4 | 4 | 5 | 4 | 4 | 4 | 4 | 4 |
uL dye | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 | 2 |
loaded | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 | 7 |
4. Ran gel for 30 minutes at 200 V. 5. Stained for 10 minutes with 3 uL SYBR gold 6. Measured concentration in Nanodrop 7. Stored at 4C for exonuclease digestion tomorrow
Nanodrop Results[edit]
Sample | ng/uL | uL | ng |
V4S3A | 102.2 | 30 | 3066 |
V4S3B | 86.8 | 30 | 2604 |
V6S3A | 80.1 | 30 | 2403 |
V6S3B | 77.9 | 30 | 2337 |