Difference between revisions of "Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2014/04/29"

From OpenWetWare
Jump to: navigation, search
(Autocreate 2014/04/29 Entry for Haynes_Lab:Notebook/Characterizing_AHL_quorum_sensing_homologs)
 
(mm/dd/yyyy)
Line 7: Line 7:
 
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
 
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
 
==mm/dd/yyyy==
 
==mm/dd/yyyy==
* Insert content here...
+
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.  
 +
<br>
 +
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).
 +
<br>
 +
Controls
 +
Positive control was a miniprepped plasmid at 100ng/ul concentration.
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|''''''
 +
| align="center" style="background:#f0f0f0;"|'''Volume (ul)'''
 +
| align="center" style="background:#f0f0f0;"|''''''
 +
|-
 +
| Thing||Positive ctrl||Negative ctrl
 +
|-
 +
| H2O||12||12
 +
|-
 +
| Green digest buffer||2||2
 +
|-
 +
| pTet mCh||5||5
 +
|-
 +
| DpnI||1||0
 +
|}
  
 +
<br><br>
  
 +
{| {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Thing'''
 +
| align="center" style="background:#f0f0f0;"|'''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.<br>
 +
Cleaned using zymo DNA clean and concentration-5 kit (columns from gel extraction kit). Added 150ul of DNA binding buffer to DpnI reactions.
 +
<br>
 +
Gel extracted PCR reaction 1. Saw a smaller band above, think I got the correct band.
 
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
 
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
 
|}
 
|}
  
 
__NOTOC__
 
__NOTOC__

Revision as of 13:03, 29 April 2014

Owwnotebook icon.png 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.