User:Tara K. Luckau/Notebook/Team ConGen/Entry Base: Difference between revisions
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==27 September 2010 - Ordered Primers== | ==27 September 2010 - Ordered Primers== | ||
* Ordered first set of primers | * Ordered first set of primers | ||
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* Aliquots: 1000µL into each of 5 snap-caps; 4 stored in freezer, 1 stored in fridge for immediate use | * Aliquots: 1000µL into each of 5 snap-caps; 4 stored in freezer, 1 stored in fridge for immediate use | ||
==19 October 2010 - PCR Scun2 F-R== | ==19 October 2010 - PCR Scun2 F-R== | ||
* [[Image:Luckau_PCR20101019.jpg| | * [[Image:Luckau_PCR20101019.jpg|700 px]] | ||
* DNA = SCOC CPN 82 | * DNA = SCOC CPN 82 | ||
* 2 mats with lab tape, no evaporation! | * 2 mats with lab tape, no evaporation! | ||
==21 October 2010 - Gel Scun2 F-R== | ==21 October 2010 - Gel Scun2 F-R== | ||
===Make 1x TAE=== | ===Make 1x TAE=== | ||
* 20mL 50x TAE + 980 mL NanoPure | * 20mL 50x TAE + 980 mL NanoPure H<sub>2</sub>O | ||
===Cast Gel=== | ===Cast Gel=== | ||
* 100mL 1x TAE + 1.5g agarose | * 100mL 1x TAE + 1.5g agarose | ||
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* 270mL TAE + 4.05g agarose | * 270mL TAE + 4.05g agarose | ||
* (for future reference, total volume ≈ volume of gel box in cm<sup>3</sup>, ie 27cm long x 20 cm wide x 0.5 cm thick gel = 270mL of TAE) | * (for future reference, total volume ≈ volume of gel box in cm<sup>3</sup>, ie 27cm long x 20 cm wide x 0.5 cm thick gel = 270mL of TAE) | ||
==25 October 2010 - Gel Scun2 F-R== | |||
===Cast Gel=== | |||
* 270mL TAE + 4.05g agarose in 500mL Erlenmyer flask | |||
* large stirbar, hotplate set at 150, stir set at 15 for about 45 min, until clear | |||
* add 27µL GelRed | |||
* let cool in dark for about 30 min | |||
* pour into caster, set 2 24-tooth combs, cover with cardboard | |||
* let cool in dark (had to do office hours, sat for about 2 hours) | |||
==25 October 2010 - Ladder Mix== | |||
===Manufacturer specifications=== | |||
* 0.1µg ladder / mm width of well (STK at 1.0µg/µL) | |||
===Calculations=== | |||
* well width (24-tooth comb) = 6mm | |||
* 6mm x 0.1µg ladder | |||
Revision as of 12:58, 25 October 2010
Tara's Lab Notebook | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page |
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27 September 2010 - Ordered Primers
11 October 2010 - PCR Thermalcycler TestingPurpose
Results
Purpose
Results
12 October 2010 - PCR Thermalcycler TestingPurpose
Results
18 October 2010 - PCR Thermalcycler TestingPurpose
Results
18 October 2010 - Primer Rehydration, Scun2Purpose
Calculations
18 October 2010 - dNTP MixPurpose
Calculations
Wet Work
19 October 2010 - PCR Scun2 F-R21 October 2010 - Gel Scun2 F-RMake 1x TAE
Cast Gel
Next Try
25 October 2010 - Gel Scun2 F-RCast Gel
25 October 2010 - Ladder MixManufacturer specifications
Calculations
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