IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/14: Difference between revisions
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TesA cut at X, P and RBS cut at S, P. | TesA cut at X, P and RBS cut at S, P. | ||
Note: in order to ensure the viability of our PstI enzyme, we ran a control with some PstI from Haynes lab. Results are shown below. the PstI of Haynes lab had full digestion, while the iGEM PstI had a small amount of partial digestion, as made manifest by the three bands of the TesA in lane 2 of the gel. | Note: in order to ensure the viability of our PstI enzyme, we ran a control with some PstI from Haynes lab. Results are shown below. the PstI of Haynes lab had full digestion, while the iGEM PstI had a small amount of partial digestion, as made manifest by the three bands of the TesA in lane 2 of the gel. | ||
[[image:14_gel_.jpg|400px|gel]] | |||
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | |||
|- valign="top" | |||
| bgcolor=#cfcfcf | Reagent | |||
| bgcolor=#cfcfcf | RBS (μL) | |||
| bgcolor=#cfcfcf | TesA (μL) | |||
| rowspan="7" | <u>'''Lanes'''</u><br>Our PstI:<br>1. 1kb ladder<br>2. TesA (651 bp)<br> 3. RBS (2083)<br>Haynes PstI:<br> 4. TesA (651 bp)<br> 5. RBS (2083 bp)<br> 6. Repeat of 5 <br> | |||
| rowspan="7" | [[image:14_gel_.jpg|400px|gel]]<br> [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |||
| DNA(plasmid) || 10.0 || 10.0 | |||
|- | |||
| 10X FD buffer || 2.0 || 2.0 | |||
|- | |||
| Enzyme # 1 || SpeI: 1.0 || XbaI: 1.0 | |||
|- | |||
| PstI || 1.0 || 1.0 | |||
|- | |||
| dH<sub>2</sub>O || 6.0 || 6.0 | |||
|- | |||
|| || 20 μL @ 42°C/ ~1 Hr. | |||
|} | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | ||
Revision as of 11:01, 15 July 2014
 Project name
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Entry title
1 L of distilled water, 35 grams of LB agar powder. Autoclave for 30 minutes.
TesA cut at X, P and RBS cut at S, P. Note: in order to ensure the viability of our PstI enzyme, we ran a control with some PstI from Haynes lab. Results are shown below. the PstI of Haynes lab had full digestion, while the iGEM PstI had a small amount of partial digestion, as made manifest by the three bands of the TesA in lane 2 of the gel.
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