IGEM:Paris Bettencourt 2012/Notebooks/suicide group/day by day//2012/09/07

From OpenWetWare

Jump to: navigation, search
(Entry title)
Current revision (16:51, 22 September 2012) (view source)
(Colicin Assay)
 
Line 8: Line 8:
==Colicin Assay==
==Colicin Assay==
 +
We tested the response of ColE2 immunity in different cell stains (NEB, ZI) to Colicin E2 cells with and without IPTG.
[[Image:SGdNEB08C2060912.jpg |300px]]
[[Image:SGdNEB08C2060912.jpg |300px]]

Current revision

Project name Main project page
Previous entry      Next entry

Colicin Assay

We tested the response of ColE2 immunity in different cell stains (NEB, ZI) to Colicin E2 cells with and without IPTG.

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

Image:070912.png

Personal tools