Daniel:Notebook/HiResChrPaint/2013-6-4: Difference between revisions
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[[Daniel:Notebook/HiResChrPaint|Back to Calendar]] | [[Daniel:Notebook/HiResChrPaint|Back to Calendar]] | ||
==DpnII Digest (from [[Daniel:Notebook/HiResChrPaint|6/3/2013]])== | ==DpnII Digest (from [[Daniel:Notebook/HiResChrPaint/2013-6-3|6/3/2013]])== | ||
4. [[Daniel:Protocols/Qiaquick_Column|Column Purification]], elute with 30 uL | 4. [[Daniel:Protocols/Qiaquick_Column|Column Purification]], elute with 30 uL | ||
Line 70: | Line 70: | ||
| V6S2-A | | V6S2-A | ||
| V4S2-A* | | V4S2-A* | ||
| V4S2- | | V4S2-A* | ||
| 10 bp ladder | | 10 bp ladder | ||
| V6S2-B** | | V6S2-B** | ||
| V4S2- | | V4S2-B | ||
| V6S2- | | V6S2-B | ||
| V4S2- | | V4S2-B* | ||
| V4S2-B* | | V4S2-B* | ||
|} | |} | ||
Notes: *denotes without aa-dUTP; **denotes undigested probe | |||
2. Ran gel for 25 minutes at 250 V | 2. Ran gel for 25 minutes at 250 V | ||
3. Stained with 3 uL SYBR gold for 3 minutes | 3. Stained with 3 uL SYBR gold for 3 minutes | ||
===Gel Image=== | ===Gel Image=== | ||
[[Image:DpnDigest_060413.jpg|600px]] | |||
===Discussion=== | |||
There doesn't seem to be a difference between the 16 hour digest and the 2 hour digest, even looking only at the normal group (no aa-dUTP). | |||
Therefore tomorrow I will do the rest of the reactions for 2 hours. |
Latest revision as of 22:38, 5 June 2013
Probe Prep 3 (Started 5/29/2013)[edit]
DpnII Digest (from 6/3/2013)[edit]
4. Column Purification, elute with 30 uL 5. Measure concentrations in Nanodrop
Nanodrop Results[edit]
Sample | ng/uL | ug in 30 uL | Yield (%) |
alpha-4B | 127.1 | 3.8 | 80.9 |
alpha-6B | 122.4 | 3.7 | 78.6 |
beta-4B | 122.6 | 3.7 | 83.6 |
beta-6B | 112.5 | 3.4 | 73.3 |
TBE Gel[edit]
1. Gel setup
Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
Sample | 10 bp ladder | V4S2-B** | V4S2-A | V6S2-A | V4S2-A* | V4S2-A* | 10 bp ladder | V6S2-B** | V4S2-B | V6S2-B | V4S2-B* | V4S2-B* |
Notes: *denotes without aa-dUTP; **denotes undigested probe
2. Ran gel for 25 minutes at 250 V 3. Stained with 3 uL SYBR gold for 3 minutes
Gel Image[edit]
Discussion[edit]
There doesn't seem to be a difference between the 16 hour digest and the 2 hour digest, even looking only at the normal group (no aa-dUTP). Therefore tomorrow I will do the rest of the reactions for 2 hours.