Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/04/02: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
|||
Line 1: | Line 1: | ||
{|{{table}} width="800" | {|{{table}} width="800" | ||
|- | |- | ||
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">gBlock (DBN001) & DBN002 PCR Amplification</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">gBlock (DBN001) & DBN002 PCR Amplification; pSB1A3-GFP Restriction Digest</span> | ||
|style="background-color: #F2F2F2" 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>}} | |style="background-color: #F2F2F2" 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>}} | ||
|- | |- |
Revision as of 11:45, 22 April 2015
gBlock (DBN001) & DBN002 PCR Amplification; pSB1A3-GFP Restriction Digest | <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> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
OverviewRunning out of the gBlock ordered (DBN001), and it's not yet in a plasmid, so I'm making more by PCR amplification. gBlock PCR run details:
Run is saved as DBN20140402B on the Bio-Rad thermal cycler. DBN002 PCR Run
Run is saved as DBN20140402A on the Bio-Rad thermal cycler. pSB1A3-GFP plasmid digestion
Mix and incubate at 37°C for 15 minutes. Then heat-inactivate by incubating at 80°C for 20 minutes.
Gel Electrophoresis of DBN001, DBN002 and pSB1A3-GFP ProductsGel layout:
It looks like I ran the gel for too long, so the expected size fragment for the gBlock would have run into the pSB1A3-GFP lane. Will need to run that again. It also looks as though the pSB1A3-GFP digestion was not successful. Will need to re-try with more plasmid, as I used most of it on transfecting the cells. |