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>
|
mm/dd/yyyy
Ran PCR products from yesterday on a gel. 5ul of reactions 2-9, all 30 of 1 so I can gel extract because there should be two bands and I only want one.
DpnI digest PCR reactions 2-7 and 9 with positive and negative controls (didn't run reaction 8 because I checked the gel early and saw that there was no DNA there).
Controls
Positive control was a miniprepped plasmid at 100ng/ul concentration.
| '
|
Volume (ul)
|
'
|
| Thing |
Positive ctrl |
Negative ctrl
|
| H2O |
12 |
12
|
| Green digest buffer |
2 |
2
|
| pTet mCh |
5 |
5
|
| DpnI |
1 |
0
|
| Thing
|
Volume (ul)
|
| Digest buffer |
4
|
| PCR rxn |
25
|
| H2O |
1
|
| DpnI |
1
|
Incubated at 37°C for 55 minutes, sat on the bench for 3 minutes, heat inactivated at 80 for 5 minutes.
Cleaned using zymo DNA clean and concentration-5 kit (columns from gel extraction kit). Added 150ul of DNA binding buffer to DpnI reactions.
Gel extracted PCR reaction 1. Saw a smaller band above, think I got the correct band.
| PCR reaction
|
Description
|
'
|
260
|
280
|
260/280
|
ng/µL
|
| 1 |
Rec bb |
Gel extracted |
0.02 |
0.01 |
2 |
20.289
|
| 2 |
pLux-GFP |
DpnI digested, column purified |
0.047 |
0.024 |
2 |
47.148
|
| 3 |
pRhl-GFP |
DpnI digested, column purified |
0.05 |
0.025 |
2.004 |
50.466
|
| 4 |
pEsa-GFP |
DpnI digested, column purified |
0.032 |
0.015 |
2.117 |
32.1
|
| 5 |
pLac-LuxR |
DpnI digested, column purified |
0.046 |
0.024 |
1.935 |
45.771
|
| 6 |
pLac-RhlR |
DpnI digested, column purified |
0.06 |
0.035 |
1.692 |
59.88
|
| 7 |
pLac-RhlR |
DpnI digested, column purified |
0.041 |
0.019 |
2.2 |
41.438
|
| 9 |
pLac-EsaR |
DpnI digested, column purified |
0.038 |
0.018 |
2.135 |
37.847
|
|
|
Didn't have enough of the backbone so started another PCR. 1-4 are int receiver vector P112 P123, 5-6 are purified PCR product of receiver backbone P112 P123.
|
Project name
