Minnesota/25 July 2008

From 2008.igem.org

(Difference between revisions)
Line 11: Line 11:
|-
|-
|'''2. Redoing ligation:''' Using left over double digest from 07-24-08, But increase the amount of ligase in the reaction and performing the ligations @ 60C which is the optimal temperature for the ligase.  
|'''2. Redoing ligation:''' Using left over double digest from 07-24-08, But increase the amount of ligase in the reaction and performing the ligations @ 60C which is the optimal temperature for the ligase.  
 +
 +
{|border="1" align="left"
 +
|-
 +
!| Parts !! 10x Buffer !! H20 !! Base Vector !! Insert DNA !! T4 DNA Ligase !! Total Volume
 +
|-
 +
| BV + Tet1 || 4.0uL || 11.5uL || 3.5uL || 17.0uL || 4.0uL || 40.0uL
 +
|-
 +
| BV + Lac2 || 4.0uL || 13.5uL || 3.5uL || 15.0uL || 4.0uL || 40.0uL
 +
|-
 +
| BV + Lac/LAMBDA || 4.0uL || 15.5uL || 3.5uL || 13.0uL || 4.0uL || 40.0uL
 +
|-
 +
| BV + Tet/p22 || 4.0uL || 11.5uL ||3.5uL || 17.5uL || 4.0uL || 40.0uL
 +
|}
 +
|-
|-
|'''3. Redoing transformations:''' Redo with new ligated products by adding much less DNA/plasmid to the TOP10 Cells, b/c if put too much ligated DNA then the cells do not transform properly (as stated by iGEM troubleshooting).
|'''3. Redoing transformations:''' Redo with new ligated products by adding much less DNA/plasmid to the TOP10 Cells, b/c if put too much ligated DNA then the cells do not transform properly (as stated by iGEM troubleshooting).

Revision as of 15:23, 25 July 2008

Back to Notebook Home
Go to Previous Day (July 24)Go to Next Day (July 28)
1. No growth on plates: Experiments on 07-24-08 failed to show growth on Kanamycin resistant plates.
2. Redoing ligation: Using left over double digest from 07-24-08, But increase the amount of ligase in the reaction and performing the ligations @ 60C which is the optimal temperature for the ligase.
Parts 10x Buffer H20 Base Vector Insert DNA T4 DNA Ligase Total Volume
BV + Tet1 4.0uL 11.5uL 3.5uL 17.0uL 4.0uL 40.0uL
BV + Lac2 4.0uL 13.5uL 3.5uL 15.0uL 4.0uL 40.0uL
BV + Lac/LAMBDA 4.0uL 15.5uL 3.5uL 13.0uL 4.0uL 40.0uL
BV + Tet/p22 4.0uL 11.5uL 3.5uL 17.5uL 4.0uL 40.0uL
3. Redoing transformations: Redo with new ligated products by adding much less DNA/plasmid to the TOP10 Cells, b/c if put too much ligated DNA then the cells do not transform properly (as stated by iGEM troubleshooting).
Retrieved from "http://2008.igem.org/Minnesota/25_July_2008"