Difference between revisions of "Team:UCLA/Notebook/Spider Silk Genetics/11 August 2015"

(→‎Results)
(→‎08/11/2015)
 
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== ICA M1-15==
 
== ICA M1-15==
 +
 +
Same as previous protocol
 +
 +
PCR amplified eluted ICA product
 +
 +
{| class="wikitable"
 +
| Q5 buffer
 +
| 10 uL
 +
|-
 +
| F-03 Primer
 +
| 2.5 uL
 +
|-
 +
| G-03 Primer
 +
| 2.5 uL
 +
|-
 +
| Template
 +
| 1 uL
 +
|-
 +
| ddH2O
 +
| 32.5 uL
 +
|-
 +
| Q5
 +
| 0.5 uL
 +
|}
 +
 +
{| class="wikitable"
 +
| 98C
 +
| 30 sec
 +
|-
 +
| 98C
 +
| 10 sec
 +
|-
 +
| 66C
 +
| 20 sec
 +
|-
 +
| 72C
 +
| 50 sec
 +
|-
 +
|
 +
| x25
 +
|-
 +
| 72C
 +
| Hold
 +
|-
 +
| 12C
 +
| Hold
 +
|}
  
 
==Results==
 
==Results==
  
 
[[File:BsaIDig0811.jpg|800px]]
 
[[File:BsaIDig0811.jpg|800px]]
 +
 +
* Lane 1 and 2
 +
** Brightest band was not the right size
 +
** Band of expected size could barely be seen in lane 1 but not in lane 2
 +
** Might have to redo ICA
 +
 +
* All BsaI digestion samples had the right band
 +
 +
== T4 ligation of digested products from [https://2015.igem.org/Team:UCLA/Notebook/Spider_Silk_Genetics/10_August_2015 08/10/2015]==
 +
 +
Transformed with ligation products

Latest revision as of 07:07, 13 August 2015

iGEM UCLA




08/11/2015

BsaI Digestion of M2AB, BC and CA

2AB 2BC 2CA
Cutsmart 5 uL 5 uL 5 uL
Template 35 uL 19.5 uL 18.2 uL
ddH2O 6 uL 21.5 uL 22.8 uL
BsaI 4 uL 4 uL 4 uL

50 C for 2hr 65 C for 20 min

ICA M1-15

Same as previous protocol

PCR amplified eluted ICA product

Q5 buffer 10 uL
F-03 Primer 2.5 uL
G-03 Primer 2.5 uL
Template 1 uL
ddH2O 32.5 uL
Q5 0.5 uL
98C 30 sec
98C 10 sec
66C 20 sec
72C 50 sec
x25
72C Hold
12C Hold

Results

BsaIDig0811.jpg

  • Lane 1 and 2
    • Brightest band was not the right size
    • Band of expected size could barely be seen in lane 1 but not in lane 2
    • Might have to redo ICA
  • All BsaI digestion samples had the right band

T4 ligation of digested products from 08/10/2015

Transformed with ligation products