User:Robwarden/Notebook/Caspase-3 Activatable Probe/2009/09/10: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 24: Line 24:
|
|
{| border="1" style="text-align:center"
{| border="1" style="text-align:center"
|+ Gel 1
|+ '''Gel 1'''
!Lane
!Lane
|1 || 2 || 3 || 4 || 5 || 6 || 7 || 8 || 9 || 10 || 11 || 12 || 13 || 14 || 15
|1 || 2 || 3 || 4 || 5 || 6 || 7 || 8 || 9 || 10 || 11 || 12 || 13 || 14 || 15
Line 35: Line 35:


{| border="1" style="text-align:center"
{| border="1" style="text-align:center"
|+ Gel 2
|+ '''Gel 2'''
!Lane
!Lane
|1 || 2 || 3 || 4 || 5 || 6 || 7 || 8 || 9 || 10 || 11 || 12 || 13 || 14 || 15
|1 || 2 || 3 || 4 || 5 || 6 || 7 || 8 || 9 || 10 || 11 || 12 || 13 || 14 || 15
Line 48: Line 48:
**Incorrect: 6297, 1434
**Incorrect: 6297, 1434


{{Image gallery
{|
|title=Gel Images
|+ '''Gel Images'''
|lines=1
|[[Image:091009-hgLucColonies1to10.jpg|thumb|Gel 1]]
|091009-hgLucColonies1to10.jpg|Gel 1|Gel 1
|[[Image:091009-hgLucColonies11to18.jpg|thumb|Gel 2]]
|091009-hgLucColonies11to18.jpg|Gel 2|Gel 2
|}
}}
 
*All colonies appeared to work, except for #12
 
==Sequencing==
Colonies 2, 5, 7, and 13 were submitted for sequencing.  T7 was used for the forward primer. BHG was used for the reverse primer.  Both primers were supplied by the sequencing facility.




Revision as of 12:23, 10 September 2009

Caspase-3 Activatable Probe <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Colony Screening

Miniprep

with Qiagen kit

Digestion

  • 20 μL reaction volume
  • NEB Buffer 4
  • .5 μL of EcoRI
  • .5 μL of SgrAI
  • 10 μL of DNA
  • Incubate at 37°C for 1 hr

Agarose Gel

  • 1% Agarose, run for 30 min at 120V
Gel 1
Lane 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Sample - 1kb 1 2 3 4 5 6 7 8 9 10 100bp
Gel 2
Lane 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Sample - 1kb 11 12 13 14 15 16 17 18 100bp
  • Predicted Banding Patterns
    • Correct: 6535
    • Incorrect: 6297, 1434
Gel Images
Gel 1
Gel 2
  • All colonies appeared to work, except for #12

Sequencing

Colonies 2, 5, 7, and 13 were submitted for sequencing. T7 was used for the forward primer. BHG was used for the reverse primer. Both primers were supplied by the sequencing facility.



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Robwarden/Notebook/Caspase-3_Activatable_Probe/2009/09/10&oldid=348889"