IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/14

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Entry title)
(Entry title)
Line 17: Line 17:
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
 +
|-
 +
|| &nbsp;  || 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 14:01, 15 July 2014

Project name Main project page
Previous entry      Next entry

Entry title

  • Made LB agar plates

1 L of distilled water, 35 grams of LB agar powder. Autoclave for 30 minutes.

  • Ran Digestion of TesA and RBS

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.


Reagent RBS (μL) TesA (μL) Lanes
Our PstI:
1. 1kb ladder
2. TesA (651 bp)
3. RBS (2083)
Haynes PstI:
4. TesA (651 bp)
5. RBS (2083 bp)
6. Repeat of 5
gel
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
dH2O 6.0 6.0
  20 μL @ 42°C/ ~1 Hr.


Personal tools