User:Karmella Haynes/Notebook/PcTF Genomics/2015/03/20: Difference between revisions

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* PAGE - glucagon & insulin (for Gbox demo next week)
* PAGE - glucagon & insulin (for Gbox demo next week)


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* Loading vol. 20 uL = 5.0 μL 4x Novagen sample buffer (blue) + 1.0 μL 1M DTT + 14 uL protein
* Loading vol. 20 uL = 5.0 μL 4x Novagen sample buffer (blue) + 1.0 μL 1M DTT + 14 uL protein
* Dilutions - make enough for at least two lanes plus pipetting error (x3)
* Dilutions - make enough for at least two lanes plus pipetting error (x3)
{|
{| class="wikitable" border="0" cellspacing="3"
|-
|-
| Protein | Final mass | Stock μL (x3) | dH<sub>2</sub>O μL (x3)
| Protein | Final mass | Stock μL (x3) | dH<sub>2</sub>O μL (x3)
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|-
|-
| Glucagon | 0.1 μg | 0.1 (0.3) | 13.9 (41.7)  
| Glucagon | 0.1 μg | 0.1 (0.3) | 13.9 (41.7)  
|-
| Insulin* | 10.0 μg | 10.0 (30.0) | 4.0 (12.0)
|-
| Insulin* | 1.0 μg | 1.0 (3.0) | 13.0 (39.0)
|-
| Insulin* | 0.1 μg | 0.1 (0.3) | 13.9 (41.7)
|}
|}
*For insulin, start with a 1/25 dilution of the 25 mg/mL stock
* 4x sample buffer batch (x15) = 75.0 4x sample buffer + 15.0 1M DTT




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# Page Ruler plus (15 uL)
# Page Ruler plus (15 uL)
# Glucagon 10.0 μg
# Glucagon 10.0 μg
# Glucagon 1.0 μg
# Glucagon 0.1 μg
# Insulin 10.0 μg
# Insulin 1.0 μg
# Insulin 0.1 μg
# Page Ruler plus (15 uL)
# Glucagon 10.0 μg
# Glucagon 1.0 μg
# Insulin 10.0 μg
# Insulin 1.0 μg




Revision as of 12:31, 23 March 2015

Pc-TF Genomics <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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03/20/15

  • PAGE - glucagon & insulin (for Gbox demo next week)


PAGE - glucagon & insulin
> From David Barclay's pancreas cell project


Sample prep

  • Loading vol. 20 uL = 5.0 μL 4x Novagen sample buffer (blue) + 1.0 μL 1M DTT + 14 uL protein
  • Dilutions - make enough for at least two lanes plus pipetting error (x3)
Final mass | Stock μL (x3) | dH2O μL (x3)
10.0 μg | 10.0 (30.0) | 4.0 (12.0)
1.0 μg | 1.0 (3.0) | 13.0 (39.0)
0.1 μg | 0.1 (0.3) | 13.9 (41.7)
10.0 μg | 10.0 (30.0) | 4.0 (12.0)
1.0 μg | 1.0 (3.0) | 13.0 (39.0)
0.1 μg | 0.1 (0.3) | 13.9 (41.7)
  • For insulin, start with a 1/25 dilution of the 25 mg/mL stock
  • 4x sample buffer batch (x15) = 75.0 4x sample buffer + 15.0 1M DTT


> Gel lanes (for both gels)

  1. Page Ruler plus (15 uL)
  2. Glucagon 10.0 μg
  3. Glucagon 1.0 μg
  4. Glucagon 0.1 μg
  5. Insulin 10.0 μg
  6. Insulin 1.0 μg
  7. Insulin 0.1 μg
  8. Page Ruler plus (15 uL)
  9. Glucagon 10.0 μg
  10. Glucagon 1.0 μg
  11. Insulin 10.0 μg
  12. Insulin 1.0 μg


Reagent Volume
reagent 1 # μL
reagent 2 #
reagent 3 #
reagent 4 #
dH2O #
  # μL

--> Reaction conditions



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