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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* Have: Invitrogen 100mM dNTP Set (catalog# 10297-018) (250µL of 25µmol in H2O @ 100mM each) | * Have: Invitrogen 100mM dNTP Set (catalog# 10297-018) (250µL of 25µmol in H2O @ 100mM each) | ||
* Want: 10mM dNTP mix (2.5mM each), 5 aliquots of 1000µL | * Want: 10mM dNTP mix (2.5mM each), 5 aliquots of 1000µL | ||
* ( | * (C<sub>1</sub>) (V<sub>1</sub>) = (C<sub>2</sub>) (V<sub>2</sub>) | ||
*: (2.5mM)(5000µL)=(100mM)( | *: (2.5mM) (5000µL) = (100mM) (V<sub>2</sub>) | ||
*: | *: V<sub>2</sub> = 125µL of each dNTP | ||
===Wet Work=== | ===Wet Work=== | ||
* 125µL dATP + 125µL dTTP + 125µL dCTP + 125µL dGTP + 4500µL H2O = 5000µL dNTP Mix @ 10mM (2.5mM each dNTP) | * 125µL dATP + 125µL dTTP + 125µL dCTP + 125µL dGTP + 4500µL H2O = 5000µL dNTP Mix @ 10mM (2.5mM each dNTP) | ||
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===Calculations=== | ===Calculations=== | ||
* well width (24-tooth comb) = 6mm | * well width (24-tooth comb) = 6mm | ||
* 6mm x 0.1µg ladder | * 6mm x $0.1µg ladder \over 1mm width$ x $µL \over 1.0µg$ = 0.6µL ladder per well | ||
* from Jeremy (Bohonak lab): uses 5µL PCR product + 1.5µL dye | |||
* Per well: 0.6µL ladder + 4.4µL H<sub>2</sub>O = 5µL ladder mix | |||
* (xylene and bromophenol blue) | |||
Revision as of 14:00, 25 October 2010
Tara's Lab Notebook | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page |
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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