Difference between revisions of "IGEM:Paris Bettencourt 2012/Notebooks/suicide group/day by day//2012/09/07"

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|}
 
|}
 +
 +
==Biobricks==
 +
We put pSG.008 (pLac+ColE2 immunity) and pSG.014 (ColE2 immunity) inside Biobricks vector (pSB1C3). All of them were cut with XbaI and PstI.
 +
===Protocol===
 +
 +
{| style="border-spacing:0;"
 +
| style="border-top:0.002cm solid #000000;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"|
 +
| style="border-top:0.002cm solid #000000;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| PSB1C3 (-)
 +
| style="border-top:0.002cm solid #000000;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| BB+pSG.008
 +
| style="border:0.002cm solid #000000;padding:0.097cm;"| BB+pSG.014
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| H2O
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 6.5 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| -
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:0.002cm solid #000000;padding:0.097cm;"| -
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| Insert
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 2 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 2 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:0.002cm solid #000000;padding:0.097cm;"| 2 uL
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| Vector
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| -
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 6.5 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:0.002cm solid #000000;padding:0.097cm;"| 6.5 uL
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| Ligase buffer 10x
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 1.0 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 1.0 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:0.002cm solid #000000;padding:0.097cm;"| 1.0 uL
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| T4 ligase enzyme
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 0.5 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 0.5 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:0.002cm solid #000000;padding:0.097cm;"| 0.5 uL
 +
 +
|-
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| Total
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 10 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:none;padding:0.097cm;"| 10 uL
 +
| style="border-top:none;border-bottom:0.002cm solid #000000;border-left:0.002cm solid #000000;border-right:0.002cm solid #000000;padding:0.097cm;"| 10 uL
 +
 +
|}
 +
 +
===Transformation===
 +
We transformed 5 uL of each ligation product with heat shock transformation; +pSG.001 as a positive control
 +
 +
===Gel===
 +
We ran the rest of the ligation product + 3 uL of green buffer; in a gel
 +
[[Image:070912.png]]
  
 
__NOTOC__
 
__NOTOC__
 
[[category:OWWLabNotebookV1]]
 
[[category:OWWLabNotebookV1]]

Revision as of 09:23, 7 September 2012

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Entry title

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SGdNEB08C2060912.jpg SGNEB08+C2060912.jpg SGdNEB08+C2060912.jpg

SGNEB0823C2060912.jpg SGNEB0823+C2060912.jpg SGdNEB0823+C2060912.jpg

SGZ108C2060912.jpg SGZ108+C2060912.jpg SGdZ108+C2060912.jpg

SGZ10823C2060912.jpg SGZ10823+C2060912.jpg SGdZ10823+C2060912.jpg

SGWTWT023060912.jpg SGWTC2060912.jpg SGWT023C2060912.jpg

Biobricks

We put pSG.008 (pLac+ColE2 immunity) and pSG.014 (ColE2 immunity) inside Biobricks vector (pSB1C3). All of them were cut with XbaI and PstI.

Protocol

PSB1C3 (-) BB+pSG.008 BB+pSG.014
H2O 6.5 uL - -
Insert 2 uL 2 uL 2 uL
Vector - 6.5 uL 6.5 uL
Ligase buffer 10x 1.0 uL 1.0 uL 1.0 uL
T4 ligase enzyme 0.5 uL 0.5 uL 0.5 uL
Total 10 uL 10 uL 10 uL

Transformation

We transformed 5 uL of each ligation product with heat shock transformation; +pSG.001 as a positive control

Gel

We ran the rest of the ligation product + 3 uL of green buffer; in a gel 070912.png