Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

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Joined: Sun Sep 25, 2005 5:10 am


Post by lara » Tue Jun 10, 2008 11:44 am

Now this has been puzzling me for sometime.The following is from the
paper titled "Quorum-sensing system influences root colonization and
biological control
ability in Pseudomonas fluorescens 2P24"
(Hai-Lei Wei,and Li-Qun Zhang).can anyone help me out with this? the
trouble is can't visualize it and without it can't proceed.....

To create a pcoI gene deletion allele, two fragments
flanking pcoI gene were amplified by PCR. One was created by primers
(5'-TCGTTCGCTGCAGGAAACC-3') which included the Pst1 site and I2056 (5'-
ATGGATCCCGGAGTTTGTGCATGCCC-3')which introduced the BamH1 site and the
other was created by primers I2328 (5'-
AAGGATCCGGGTTCGGCTTCTCTGACAC-3') which introduced the BamH1 site and
3') which introduced the EcoR1 site.The standard PCR was carried
out.After being digested with relevant restriction enzymes, the two
fragments were inserted into pBSNot6.

King Cobra
King Cobra
Posts: 635
Joined: Thu Feb 14, 2008 7:40 pm

Re: pcr

Post by Cat » Tue Jun 10, 2008 10:14 pm

Normal pcoI gene:

-- Flanking region 1 -- pcoI gene -- Flanking region 2 ---

Deletion of pcoI gene:

-(Pst1)- Flanking region 1 -- (BamH1) -- Flanking region 2 -(EcoR1)--

Homologous recombination substitutes normal allele with the deletion.

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