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A lot of plant genomes are big- wheat for example is 12 Gb, so it may not be quite as trivial as one might expect. cheers, tom Tom Peat Proteins Group Biomedical Program, CSIRO 343 Royal Parade Parkville, VIC, 3052 +613 9662 7304 +614 57 539 419 [log in to unmask] ________________________________________ From: CCP4 bulletin board <[log in to unmask]> on behalf of Keller, Jacob <[log in to unmask]> Sent: Tuesday, July 25, 2017 1:43 AM To: [log in to unmask] Subject: Re: [ccp4bb] Primer design >No need of the whole exome. Sequencing The second PCR product will do the job I guess. Second PCR (from the cDNA pool) with specific forward primer and and oligodA reverse primer. Surely a matter of less than $3 $3 is a major understimation, but I see your point. On the other hand, it is important to consider new technologies, and it would be a service to the scientific community to publish the exome somewhere, so other researchers would not have to spend their $3. JPK Best, DKG ----- Original Message ----- From: "Jacob Keller" <[log in to unmask]> To: "Debasish Kumar Ghosh" <[log in to unmask]>, [log in to unmask] Sent: Monday, July 24, 2017 6:48:25 PM Subject: RE: Primer design ....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 oligodA 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