From 2008.igem.org
(Difference between revisions)
|
|
Line 14: |
Line 14: |
| | | |
| | | |
- | <li>Digestion of nazwa (Gfp genetrator), nazwa (RFP generator) with NotI</li> | + | <li>Digest of nazwa (Gfp genetrator), nazwa (RFP generator) with NotI</li> |
| | | |
| <li> | | <li> |
- | Digestion of pZC with NotI and defosforylation with CIAP
| + | Digest of pZC with NotI and defosforylation with CIAP |
| </li> | | </li> |
| | | |
Revision as of 09:22, 16 October 2008
|
|
|
|
Change of the reporter from pZC with B-galactosidaze to GFP or RFP
Piotr, Weronika
- Isolation of nazwa (Gfp genetrator), nazwa (RFP generator)
- Digest of nazwa (Gfp genetrator), nazwa (RFP generator) with NotI
-
Digest of pZC with NotI and defosforylation with CIAP
-
Gel electrophoresis and gel out of proper bands
-
Ligation of pZC with nazwa (Gfp genetrator), nazwa (RFP generator)
- Chemotransformation of E.coli TOP10 with ligation products
- Plating transformants on LB+Amp30+X-gal+IPTG
Preparation of constructs with OmpA protein fusionsMichał K.
Clean-up of OmpA_alpha and OmpA_omega digest products.
Cloning PCL products on pKS vector
Michał L., Ewa, Marcin
- Gel electophoresis of PCL products (Fig. 1).
- Gel-out of proper bands (alpha-A: 1500 bp and omega-A: 1440 bp).
- Restriction digest of pKS vector and the PCL products with NotI and SacI (BamHI buffer) for 2 hours.
- Ligation 1 hour.
- Transformation of Top10 strain and screening on Amp100 plates.
Fig. 1.PCL product - Omega-A fusion.
|
|