Dear Syed,

The process is very trivial to clone your gene of interest. Assuming your gene 
is transcribed as mono-cistronic mRNA, take the oligodT primer as reverse 
primer. First isolate the total RNA from the tissue or cells and do the cDNA 
synthesis using oligodT primer followed by gene specific PCR with forward 
primer and oligodT primer from the cDNA pool. Sequence the PCR product to get 
to know the first stop codon in the ORF. Making the specific reverse primer 
from the sequence is then just matter of time.  
Hope this helps.

Best wishes,

Debasish

CSIR- Senior Research Fellow (PhD Scholar)
C/o: Dr. Akash Ranjan
Computational and Functional Genomics Group
Centre for DNA Fingerprinting and Diagnostics
Hyderabad, INDIA

Email(s): dkgh...@cdfd.org.in, dgho...@gmail.com
Telephone: 0091-9088334375 (M), 0091-40-24749396 (Lab)
Lab URL: http://www.cdfd.org.in/labpages/computational_functional_genomics.html



----- Original Message -----
From: "syed ibrahim" <0000048c02cac012-dmarc-requ...@jiscmail.ac.uk>
To: CCP4BB@JISCMAIL.AC.UK
Sent: Monday, July 24, 2017 4:56:00 PM
Subject: [ccp4bb] Primer design

Hello All

I am interested in cloning a gene from a plant. I searched the database only 
partial sequence is available, ie: for 160 residues only. The full length of 
the protein is around 570 residues. I designed forward primer and I have no 
clue to design reverse primer.

Any help

Thank you

Syed

Reply via email to