Haynes Lab:Notebook/Protein-DNA fusions in living microbes/2013/10/09

From OpenWetWare
Revision as of 22:58, 2 December 2013 by Ryan Muller (talk | contribs) (Entry title)
Jump to: navigation, search
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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Entry title

Performed successful tranformation with Kylie's BL21(DE3) used protocol here: http://www.molecularinfo.com/MTM/A/A3/A3-1.pdf

Made SOC media: LB miler broth + tryptone + 20 mM MgSO4 and 2.5 mM KCl (http://openwetware.org/wiki/SOB)

Grew up 4 colonies of BL21(DE3) in Kan media, used 3 of them (1, 2, 4) to induce expression of Topo per protocol: http://www.molecularinfo.com/MTM/A/A3/A3-1.pdf

Ran PCR using 1-omega F TSE and alpha-4 R SP and PSV plasmid template. PCR product has 3 topo cut sites.

Ran gel of wheat-germ topo +RFP pSB1C3 plasmid. From left to right: 2uL, 1.5uL, 1uL, 0.5uL, blank glycerol stock (15/30 uL in each well)

Made competent cell stock of BL21(DE3) from cells given by Kylie

Used Bug buster on induced samples (1, 2, 4) per protocol: http://www.ebiotrade.com/buyf/productsf/novagen/70584-000.pdf