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

From OpenWetWare
Jump to navigationJump to search
(fix raw html notebook nav)
 
(3 intermediate revisions by one other user not shown)
Line 2: Line 2:
|-
|-
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Pc-TF Genomics</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
|-
|-
| colspan="2"|
| colspan="2"|
Line 11: Line 11:
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
* PAGE - glucagon & insulin (for Gbox demo next week)
* PAGE - glucagon & insulin (for Gbox demo next week)


----
----
Line 20: Line 21:
* 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)
|-
| Glucagon || 10.0 μg || 10.0 (30.0) || 4.0 (12.0)
|-
| Glucagon || 1.0 μg || 1.0 (3.0) || 13.0 (39.0)
|-
|-
| Protein | Final mass | Stock μL (x3) | dH<sub>2</sub>O μL (x3)
| Glucagon || 0.1 μg || 0.1 (0.3) || 13.9 (41.7)  
|-
|-
| Glucagon | 10.0 μg | 10.0 (30.0) | 4.0 (12.0)  
| Insulin* || 10.0 μg || 10.0 (30.0) || 4.0 (12.0)  
|-
|-
| Glucagon | 1.0 μg | 1.0 (3.0) | 13.0 (39.0)  
| Insulin* || 1.0 μg || 1.0 (3.0) || 13.0 (39.0)  
|-
|-
| Glucagon | 0.1 μg | 0.1 (0.3) | 13.9 (41.7)  
| 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




Line 35: Line 46:
# 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




Latest revision as of 00:50, 27 September 2017

Pc-TF Genomics Main project page
Previous entry      Next entry


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)
Protein Final mass Stock μL (x3) dH2O μL (x3)
Glucagon 10.0 μg 10.0 (30.0) 4.0 (12.0)
Glucagon 1.0 μg 1.0 (3.0) 13.0 (39.0)
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


> 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



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Karmella_Haynes/Notebook/PcTF_Genomics/2015/03/20&oldid=1016560"