Difference between revisions of "Team:UCLA/Notebook/Spider Silk Genetics/14 August 2015"

(→‎08/14/2015)
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| Hold  
 
| Hold  
 
|}
 
|}
 +
 +
Ran samples on 1% gel
 +
 +
Sample 1 with had no band
 +
Sample 2 had a very bright band close to the expected size
 +
 +
Maybe it was the ICA elution that was no longer in good condition?
 +
 +
Gel purified the band from sample 2
 +
* 39.69 ng/uL; 260/280: 2.47
 +
 +
==Double Digestion of M1-12 PCR product with EcoRI and PstI==
 +
 +
{| "class="wikitable"
 +
| EcoRI
 +
| 1 uL
 +
|-
 +
| Pst I
 +
| 1 uL
 +
|-
 +
| Template
 +
| 11 uL
 +
|-
 +
| Buffer 2.1
 +
| 5 uL
 +
|-
 +
| ddH2)
 +
| 32 uL
 +
|}
 +
 +
37C for 1.5 hr, 65C for 20 min

Revision as of 03:20, 15 August 2015

iGEM UCLA




08/14/2015

PCR amplify M1-12

Since previous PCRs using ICA elution as template did not work very well, first M1-12 PCR product was also tested as a template

ICA elution PCR product
Q5 buff 10 uL 10 uL
dNTPs 1 uL 1 uL
F-03 2.5 uL 2.5 uL
G-03 2.5 uL 2.5 uL
ddH2O 32.5 uL 33.3 uL
Template 1 uL 0.2 uL
Q5 Polymerase 0.5 uL 0.5 uL
98C 30 sec
98C 10 sec
66C 20 sec
x23
72C 45 sec
72C 2 min
12C Hold

Ran samples on 1% gel

Sample 1 with had no band Sample 2 had a very bright band close to the expected size

Maybe it was the ICA elution that was no longer in good condition?

Gel purified the band from sample 2

  • 39.69 ng/uL; 260/280: 2.47

Double Digestion of M1-12 PCR product with EcoRI and PstI

EcoRI 1 uL
Pst I 1 uL
Template 11 uL
Buffer 2.1 5 uL
ddH2) 32 uL

37C for 1.5 hr, 65C for 20 min