Request for sequencing a whole chromosome cell line

Iredc27 · 10-05-2011, 05:03 AM

Hi to everyone, i need your help!
I am currently working in Cytogenetics in a lab located in Montevideo (Uruguay), and i am working with a mouse cell line. I am able to identify each chromosome of this cell line, based on its unique pattern of dark and light bands. I am interested in studying the sequence of the chromosome's pair 1. Through Chromosome microdissection I physically removes the complete chromosome of pair 1 from the slide. Then i procedure to make multiple copies of the isolated Chromosome using Sigma GenomePlex Single Cell Whole Amplification Kit, that work in three step 1)fragment the chromosome dna, 2)prepare a DNA library, 3)amplification of the DNA fragment
I need to sequence the whole chromosome, but in Montevideo, we don not account even with this new and incredible technology in sequencing DNA.
I was wonderng if this amplified sample of the entire chromosome (explaind before) would work if i send it to sequence with Illumina Hiseq2000 for example?

Which other things i have to take into consideration?

I really appreciate any response, criticisms or recommendations

Ire

flxlex · 10-06-2011, 02:49 AM

Closed due to cross-posting to in total four forums.

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10-05-2011, 05:03 AM	#1
Iredc27 Junior Member Location: Montevideo, Uruguay Join Date: Oct 2011 Posts: 8	Request for sequencing a whole chromosome cell line Hi to everyone, i need your help! I am currently working in Cytogenetics in a lab located in Montevideo (Uruguay), and i am working with a mouse cell line. I am able to identify each chromosome of this cell line, based on its unique pattern of dark and light bands. I am interested in studying the sequence of the chromosome's pair 1. Through Chromosome microdissection I physically removes the complete chromosome of pair 1 from the slide. Then i procedure to make multiple copies of the isolated Chromosome using Sigma GenomePlex Single Cell Whole Amplification Kit, that work in three step 1)fragment the chromosome dna, 2)prepare a DNA library, 3)amplification of the DNA fragment I need to sequence the whole chromosome, but in Montevideo, we don not account even with this new and incredible technology in sequencing DNA. I was wonderng if this amplified sample of the entire chromosome (explaind before) would work if i send it to sequence with Illumina Hiseq2000 for example? Which other things i have to take into consideration? I really appreciate any response, criticisms or recommendations Ire __________________ Ire rolleyes:

10-06-2011, 02:49 AM	#2
flxlex Moderator Location: Oslo, Norway Join Date: Nov 2008 Posts: 415	Closed due to cross-posting to in total four forums.