Hi there!
I want to sequence the whole Y chromosome and i'm looking for the method of enrichment for it. Could anybody suggest any approach for that?
Thanks in advance!
Hi Nazar,
depending on the species, but most Y chromosomes are larger than 30 Mb... there is a paper made on gorilla (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4817776/), you could take the advices.
fred
Dear Nazar Samarov,
As Frederic already pointed out, the species information is crucial. In the old days, Y chromosomes were isolated using flow cytometry, based on their exceptional DNA content.I believe it was at one time common to sort bull sperm to avoid the ocurrence of male calves when doing artificial insemination. So, the first thing you would have to do is to determine the size of the Y chromosome versus all the others, (and if sufficiently different from the other chromosomes) synchronize cells, lyse them and sort the chromosomes. We did try something similar with plants, but without success at that time due to cloning and sequencing problems. http://onlinelibrary.wiley.com/doi/10.1002/cyto.990210408/pdf
I hope this helps with your project. Kind regards, Harrie Verhoeven.
Maybe a little provocative answer but given the funding is there why not do whole genome sequencing? There would be considerable work-hours saving versus flow sorting, clean-up and preparation and thereafter sequencing.
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