Daniel:Notebook/ComboLock/2017-1-26: Difference between revisions
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<ol start="2"> | <ol start="2"> | ||
<li>qPCR</li> | <li>qPCR</li> | ||
<ol type="A"> | |||
<li>Make 5.2X qPCR master mix according to following recipe</li> | |||
<ol type="a"> | |||
<li>109.2 uL nfH2o</li> | |||
<li>5.2 uL AmpF</li</li> | |||
<li>130 uL 2X Kapa SYBR Fast Master Mix</li> | |||
<li>Add 47 uL master mix to each well</li> | |||
<li>Add 2 uL sample and 1 uL appropriate Reverse Primer (1 uM) according to table</li> | |||
<li>qPCR Cycles</li> | |||
<ol type="a"> | |||
<li>95C 3 min</li> | |||
<li>95C 3 sec</li> | |||
<li>55C 30 sec</li> | |||
<li>72C 20 sec</li> | |||
<li>plate read</li> | |||
<li>goto b x24</li> | |||
<li>72C 2 min</li> | |||
<li>16C hold</li> | |||
</ol></ol> | |||
****** | |||
<li>TBE Gel</li> | |||
<ol type="A"> | |||
<li>Mix 64 uL TBE, 16 uL 6x loading dye</li> | |||
<li>Aliquot 10 uL per sample/ladder lane onto parafilm</li> | |||
<li>Add 2 uL of sample or ladder to correct drop</li> | |||
<li>Load 10 uL in to well</li> | |||
<li>Run gel for 23 minutes at 230V</li> | |||
<li>Open gel and stain with 3 uL SYBR Gold for 3 minutes</li> | |||
<li>Rinse gel and image in gel doc</li> | |||
</ol></ol> | |||
===Results=== | |||
<gallery perrow=2 heights=300px widths=300px> | |||
File:|qPCR curve | |||
File:|Gel image | |||
</gallery> |
Revision as of 17:01, 26 January 2017
RCA Test (Started Monday)
Epicentre Test (Started Yesterday)
Sample Matrix
- qPCR
- Make 5.2X qPCR master mix according to following recipe
- 109.2 uL nfH2o
- 5.2 uL AmpF</li
- 130 uL 2X Kapa SYBR Fast Master Mix
- Add 47 uL master mix to each well
- Add 2 uL sample and 1 uL appropriate Reverse Primer (1 uM) 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 23 minutes at 230V
- Open gel and stain with 3 uL SYBR Gold for 3 minutes
- Rinse gel and image in gel doc