Haynes Lab:Notebook/Engineering PC-TFs/2013/01/25: Difference between revisions
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(Autocreate 2013/01/25 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs) |
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== | ==01/25/13== | ||
* | * Type IIS assembly - day 1 | ||
---- | |||
PCR | |||
* Resuspend new primers to a final concentration of 100 μM | |||
* Make 10 μM dilutions in separate tubes | |||
* All of the reactions are as follows: | |||
# template, primer 1, primer 2 | |||
# template, primer 1, primer 2 | |||
# template, primer 1, primer 2 | |||
# template, primer 1, primer 2 | |||
{| {{table}} border="1" cellspacing="3" <!-- PCR rxn. table --> | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | Volume | |||
| rowspan="7" | <u>Expected:</u><br>1. pSB1A3 = ~2000<br>2. hPCD = 186<br>3. BL01 = 2520 | |||
| rowspan="7" | [[Image:KAH011013_gel1.jpg|200px|Hover name]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA(plasmid) || 0.2 μL | |||
|- | |||
| primer 1 || 1.0 | |||
|- | |||
| primer 2 || 1.0 | |||
|- | |||
| 2x GoTaq || 25.0 | |||
|- | |||
| dH<sub>2</sub>O || 22.8 | |||
|- | |||
| || 50.0 μL | |||
|} | |||
PCR | |||
* 95°C, 3 min. | |||
* [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35 | |||
* 72°C, 3 min. | |||
* 4°C, ∞ | |||
Revision as of 13:38, 25 January 2013
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01/25/13
PCR
PCR
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