Haynes Lab:Notebook/Engineering PC-TFs/2012/11/26

From OpenWetWare

< Haynes Lab:Notebook | Engineering PC-TFs | 2012 | 11(Difference between revisions)
Jump to: navigation, search
(Autocreate 2012/11/26 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs)
Current revision (20:33, 6 September 2013) (view source)
 
(One intermediate revision not shown.)
Line 1: Line 1:
{|{{table}} width="800"
{|{{table}} width="800"
|-
|-
-
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Engineering PC-TFs</span>
+
|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: #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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
+
|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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|-
|-
| 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==
==Summary==
-
*  
+
 
 +
 
 +
*'''[[User:Karmella Haynes|---Karmella]] 01:17, 26 November 2012 (EST)''':
 +
 
 +
* Digest the pSB1A2/3 plasmid (from Rene) with XbaI & SpeI to get rid of the insert
 +
 
 +
{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table -->
 +
|-valign="top"
 +
| <u>Reagent</u> || <u>Volume</u> || &nbsp;
 +
| rowspan="9" |
 +
|-
 +
| DNA (plasmid) || 20.0
 +
|-
 +
| 10x buffer || 3.0
 +
|-
 +
| XbaI || 1.0
 +
|-
 +
| SpeI || 1.0
 +
|-
 +
| dH<sub>2</sub>O || 5.0
 +
|-
 +
| &nbsp; || 30 μL --> 37°C/ ~10 min.
 +
|}
 +
 
 +
* Run entire 30 μL on a 1% gel (use big tooth comb) next to a ladder
 +
* You should see two bands. The larger band (2000 bp) is the backbone.
 +
* Cut out and gel purify this fragment (ignore the shorter fragment).
 +
* Notify Dr. Haynes when you are done. The next step is Gibson Assembly and transformation.
 +
 

Current revision



Main project page
Previous entry      Next entry

Summary

  • Digest the pSB1A2/3 plasmid (from Rene) with XbaI & SpeI to get rid of the insert
Reagent Volume  
DNA (plasmid) 20.0
10x buffer 3.0
XbaI 1.0
SpeI 1.0
dH2O 5.0
  30 μL --> 37°C/ ~10 min.
  • Run entire 30 μL on a 1% gel (use big tooth comb) next to a ladder
  • You should see two bands. The larger band (2000 bp) is the backbone.
  • Cut out and gel purify this fragment (ignore the shorter fragment).
  • Notify Dr. Haynes when you are done. The next step is Gibson Assembly and transformation.



Personal tools