Difference between revisions of "Team:UCLA/Notebook/Spider Silk Genetics/10 August 2015"

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(→‎EcoRI+PstI Digestion of M1-12)
 
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=08/10/2015=
 
=08/10/2015=
 +
 +
==Redid PCR to amplify M1-12 ICA product==
 +
 +
{| class="wikitable"
 +
| 5X Q5 buff
 +
| 10 uL
 +
|-
 +
| 10mM dNTPs
 +
| 1 uL
 +
|-
 +
| F-03
 +
| 2.5 uL
 +
|-
 +
| G-03
 +
|2.5 uL
 +
|-
 +
| Template
 +
| 1 uL
 +
|-
 +
| ddH2O
 +
| 32.5 uL
 +
|-
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| Q5
 +
| 0.5 uL
 +
|}
 +
 +
{| class="wikitable"
 +
| 98C
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| 30 sec
 +
|-
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| 98C
 +
| 10 sec
 +
|-
 +
| 66C
 +
| 20 sec
 +
|-
 +
| 72C
 +
| 40 sec
 +
|-
 +
|
 +
| x23
 +
|-
 +
| 72C
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| 2 min
 +
|-
 +
| 12 C
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| Hold
 +
|}
 +
 +
*Changed extension time to 40 sec instead of 35 sec
 +
* Used new primers from stock
 +
 +
 +
 +
Ran samples on 1% gel with 1kb ladder
 +
 +
Gel purified: 11ng/uL in 30uL
 +
 +
==EcoRI+PstI Digestion of M1-12==
 +
 +
{| class="wikitable"
 +
| EcoRI
 +
| 1 uL
 +
|-
 +
| Pst I
 +
| 1 uL
 +
|-
 +
| 2.1 buffer
 +
| 5 uL
 +
|-
 +
| DNA
 +
| 33 uL
 +
|}
 +
 +
37C for 1.5 hr
 +
65C for 20 min

Latest revision as of 01:56, 11 August 2015

iGEM UCLA




08/10/2015

Redid PCR to amplify M1-12 ICA product

5X Q5 buff 10 uL
10mM dNTPs 1 uL
F-03 2.5 uL
G-03 2.5 uL
Template 1 uL
ddH2O 32.5 uL
Q5 0.5 uL
98C 30 sec
98C 10 sec
66C 20 sec
72C 40 sec
x23
72C 2 min
12 C Hold
  • Changed extension time to 40 sec instead of 35 sec
  • Used new primers from stock


Ran samples on 1% gel with 1kb ladder

Gel purified: 11ng/uL in 30uL

EcoRI+PstI Digestion of M1-12

EcoRI 1 uL
Pst I 1 uL
2.1 buffer 5 uL
DNA 33 uL

37C for 1.5 hr 65C for 20 min