Daniel:Notebook/ComboLock/2017-1-18: Difference between revisions
Jump to navigation
Jump to search
>Djacobse |
>Djacobse |
||
Line 106: | Line 106: | ||
<li>Add 2 uL of sample or ladder to correct drop</li> | <li>Add 2 uL of sample or ladder to correct drop</li> | ||
<li>Load 10 uL in to well</li> | <li>Load 10 uL in to well</li> | ||
<li>Run gel for 22 minutes at | <li>Run gel for 22 minutes at 230V</li> | ||
<li>Open gel and stain with 3 uL SYBR Gold for 3 minutes</li> | <li>Open gel and stain with 3 uL SYBR Gold for 3 minutes</li> | ||
<li>Rinse gel and image in gel doc</li> | <li>Rinse gel and image in gel doc</li> | ||
</ol></ol> | </ol></ol> | ||
===Results=== | |||
<gallery perrow=-2 heights=300px widths=300px> | |||
File: | |||
File:2017-01-18-v3Oligos-Adapters.png|Gel Image | |||
</gallery> |
Revision as of 00:13, 19 January 2017
Version 3 Oligos (Started Thursday, Jan 13)
Product Check
Searching for the bridge product (or circular product) using the primer24/primer12 combination.
- qPCR
- Make 5.2X qPCR master mix according to following recipe
- 109.2 uL nfH2O
- 130 uL SYBR Fast
- 5.2 uL 10 uM primer24RC
- 5.2 uL 10 uM primer24RC
- Add 48 uL master mix to each well
- Add 2 uL sample according to table
- qPCR Cycles
- 95C 3 min
- 95C 3 sec
- 55C 30 sec
- 72C 20 sec
- plate read
- goto b x24
- 72C 2 min
- 16C hold
- TBE Gel
- Mix 64 uL TBE, 16 uL 6x loading dye
- Aliquot 10 uL per sample/ladder lane onto parafilm
- Add 2 uL of sample or ladder to correct drop
- Load 10 uL in to well
- Run gel for 22 minutes at 230V
- Open gel and stain with 3 uL SYBR Gold for 3 minutes
- Rinse gel and image in gel doc
Sample | Condition | Lane | Sample Vol (uL) | 2X Kapa SYBR qPCR MM | 10 uM Forward Primer | 10 uM Reverse Primer | H2O | Total Volume (uL) |
Sample 1A | Sample | A1 | 2 | 25 | 1 | 1 | 21 | 50 |
Sample 1B | Sample | A2 | 2 | 25 | 1 | 1 | 21 | 50 |
Sample 4A | No Lock Phosphate | A3 | 2 | 25 | 1 | 1 | 21 | 50 |
Sample 4B | No Lock Phosphate | A4 | 2 | 25 | 1 | 1 | 21 | 50 |
NTC | qPCR Primers Only | A5 | 2 | 25 | 1 | 1 | 23 | 50 |
Results
- 2017-01-18-v3Oligos-Adapters.png
Gel Image