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....Or sequence the whole exome for what, $500-1000?

JPK

-----Original Message-----
From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of Debasish Kumar Ghosh
Sent: Monday, July 24, 2017 7:43 AM
To: [log in to unmask]
Subject: Re: [ccp4bb] Primer design

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): [log in to unmask], [log in to unmask]
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" <[log in to unmask]>
To: [log in to unmask]
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