Haynes Lab:Notebook/Engineering PC-TFs/2013/02/01: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 6: Line 6:
| colspan="2"|
| colspan="2"|
<!-- ##### 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. ##### -->
==Summary==
==2/1/2013==
Type IIS Day 3 Assembly
 
----
 
<font size=3>'''Bacterial transformation'''</font>
<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.
* Added total volume (10.0 μL) to 50 μL chemically competent cells  (e.g., BL21) in a 2.0 mL tube.

Revision as of 21:02, 1 February 2013

Engineering PC-TFs <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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

2/1/2013

Type IIS Day 3 Assembly

Bacterial transformation

  • 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. (Accidentally for 1 min), and placed on ice.
  • Added 800 μL sterile SOC medium.
  • Grow with shaking at 37°C for 30 min.(Was confused about how to put 2ml tubes in shaker.)
  • 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. Protocol didn't specify.) Grow overnight at 37°C.
  • Check results tomorrow.



Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/Engineering_PC-TFs/2013/02/01&oldid=672550"