Haynes Lab:Notebook/Engineering PC-TFs/2013/02/14: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
|||
(One intermediate revision by the same user not shown) | |||
Line 1: | Line 1: | ||
{|{{table}} width="800" | {|{{table}} width="800" | ||
|- | |- | ||
|style="background-color: # | |style="background-color: #FFCC00"|<br>[[Image:Hayneslab3.gif|200px|center]][[Image:Asu logo 3.gif|200px|center]]<span style="font-size:22px;"><br></span> | ||
|style="background-color: # | |style="background-color: #800000" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
Line 60: | Line 60: | ||
|} | |} | ||
<font size=3>'''Bacterial transformation'''</font> | |||
* Added total volume (10.0 μL) to 50 μL chemically competent cells (e.g., BL21) in a 2.0 mL tube. | |||
* Incubated on ice for 2 min., heat shock at 42°C for exactly 45 sec., and placed on ice. | |||
* Added 800 μL sterile SOC medium. | |||
* Grow with shaking at 37°C for 30 min.(''Taped tubes to the shaker rack.'') | |||
* Pelleted the cells at top speed in a microcentrifuge for 3 min. at room temperature. | |||
* Discarded the supernatant. Resuspended the cells in 100 μL LB + antibiotic. | |||
* Plate cells on pre-warmed LB agar + antibiotic. (''I plated 100 μL of the cells. Then put beads on plates and shook.) Protocol didn't specify.'') Grow overnight at 37°C. | |||
* Check results tomorrow. | |||
There were no colonies on the plates. It's possible that the concentration of the ligation product is to low, so I will try to amplify. | |||
<!-- ##### 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. ##### --> |
Revision as of 18:27, 6 September 2013
<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> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Summary
Bacterial transformation
There were no colonies on the plates. It's possible that the concentration of the ligation product is to low, so I will try to amplify. |