Protocols for ET cloning

ET cloning method uses homologous recombination for DNA engineering in E. coli and was developed in Stewart lab at European Molecular Biology Laboratory.

This ET cloning protocol describes in detail the cloning procedures using ET recombination technique.

General steps for ET cloning:
1. design oligonucleotide primers
2. PCR and recipient plasmid DNA preparation
3. Preparation of competent bacterial cells
4. Transformation e.coli
5. Using Cre or Flp transient plasmids
6. Using Cre or Flp expressing E.coli

References:

Background information and troubleshooting on ET cloning

Genet Eng (N Y). 2000;22:77-98.

Nature Biotechnology 18, 1314 - 1317 (2000) DNA cloning by homologous recombination in Escherichia coli

Youming Zhang, Joep P.P. Muyrers, Giuseppe Testa & A. Francis Stewart

Last update 23-Jun-2005, Rating of 1 votes.

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	By chu van trung on 23-Jun-2005 I want to have protocols for designing and constructing plasmid for knockout baff gene in mouse Rating: Reply

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