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Finding a gene within an introgression

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

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Finding a gene within an introgression

Postby arnon77 » Mon May 07, 2012 12:38 pm

Hi
I have 2 Nearly Isogenic Lines (NILs) in pepper. The NILs are different in phenotype and segregate in a mendelian pattern. The introgression is few Mbp long and I need a gene!
Be aware that capsicum is poor in markers and a genomic sequence so I'm kind of stuck here.
I thought about 2 strategies:

• From an F2:NIL-1xNIL-2 population- take 2 phenotypic pools of RNA and sequence them in order to find few differential genes which might include my gene within the introgression.
• Same as previous only make a DNA pool and sequence in order to produce new markers based on polymorphism within the introgression.

Has anyone been dealing practically with such? and could recommend a short path towards a gene?
Thanx
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Postby JackBean » Mon May 07, 2012 1:00 pm

few Mbp will probably contain more than one gene.

With RNA sequencing, you will see also differences in genes not included in your introgression. Thus, the question is, are you interested only in that part or do you want all differences?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Re: Finding a gene within an introgression

Postby arnon77 » Wed May 09, 2012 8:16 am

It's my PhD thesis, so I need a gene in the shortest time=most efficient method.
and true probably there are few dozens of genes within that segment.
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Postby JackBean » Thu May 10, 2012 1:52 pm

The question stays, are you interested in what makes the lines different or in what is in the introgression?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Re: Finding a gene within an introgression

Postby arnon77 » Tue May 15, 2012 11:17 am

sorry for bein away...
I wish i could find the actual gene that is the cause of the difference.
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Postby JackBean » Tue May 15, 2012 11:25 am

then go for the mRNA profiling
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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