SNP analysis from hematocrit? - (Feb/13/2008 )
This is likely an extremely naive question, and I apologize for that in advance.
Is it possible to do any SNP analyses using hematocrit that has been spun down from EDTA blood? If so, roughly how many ml's would it usually require, and could that hemotocrit be frozen prior to testing?
Thanks in advance for any help.
Is this human blood? I'm going to assume that you are going to use all of the elements or I should say the leukocyte portion of the blood? Mature human red blood cells do not have a nucleus and therefore no DNA so you can't do SNP analysis unless you have DNA. Therefore you would have to use the white blood cell portion of the blood since white blood cells have a nucleus.
I've made cDNA from RNA isolated from blood and have isolated genomic DNA from blood using a kit and it's pretty easy - the amount of blood needed would depend on how much RNA/ DNA you would need. You could check the protocol of a kit to figure that out. I was using leukemic blood and got quite a bit of material but you won't get so much from healthy unstimulated blood.
I'm not sure about freezing the blood but I would think it might not be good. I think it might be better to extract the DNA/ RNA as soon as possible before freezing or maybe there is a freezing medium that can be used? I always tried to extract as soon as possible especially if I want RNA.
I hope this helps.