Haynes Lab:Notebook/Engineering PC-TFs/2012/11/26: Difference between revisions
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|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"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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==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> || | |||
| rowspan="9" | | |||
|- | |||
| DNA (plasmid) || 20.0 | |||
|- | |||
| 10x buffer || 3.0 | |||
|- | |||
| XbaI || 1.0 | |||
|- | |||
| SpeI || 1.0 | |||
|- | |||
| dH<sub>2</sub>O || 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. | |||
Latest revision as of 22:17, 26 September 2017
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