The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.
 Project name
|
<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> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>
|
November 14, 2012
- The DNA was extracted from the gel slices using the Zymoclean Gel DNA Recovery Kit
| Sample |
260 |
280 |
260/280 |
ng/μL
|
| H2B |
0.006 |
0.004 |
1.463 |
6.384
|
| LOV |
0.011 |
0.004 |
2.769 |
11.492
|
| pSB1A3 |
0.004 |
0.001 |
2.857 |
4.233
|
- ---Karmella 21:34, 15 November 2012 (EST): Next step - ligation & transformation
Refer to this page for a full description: http://openwetware.org/wiki/Haynes:Assembly101
- Use 20 ng of vector for each ligation = 5 μL pSB1A3
- Use 2x moles of H2B insert compared to vector: xμL H2B insert = 2 * 410 bp H2B / 2000 bp pSB1A3 * 20 ng pSB1A3 / 6.384 ng/μL H2B = 1.28 μL
- Use 2x moles of LOV insert compared to vector: xμL LOV insert = 2 * 461 bp LOV / 2000 bp pSB1A3 * 20 ng pSB1A3 / 11.492 ng/μL LOV = 0.8 μL
| |
H2B |
LOV |
Negative Control
|
| Insert DNA (X ng) |
1.0 μL |
1.0 |
0
|
| Vector DNA (50 ng) |
5.0 |
5.0 |
5.0
|
| 2x Roche Rapid Ligation buffer |
6.0 μl |
6.0 |
6.0
|
| New England Biolabs T4 ligase |
1.0 |
1.0 |
1.0
|
| dH2O |
___ μL |
___ μL + Insert μL
|
| TOTAL |
12.0 |
12.0 |
12.0
|
Mix the reaction(s) thoroughly by flicking the tube.
Incubate at room temperature for 10 minutes.
|
|
Project name
