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RNA/cell accumulation of difficult-to-obtain tissue


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#1 cnatstop

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Posted 16 January 2011 - 01:12 PM

hello microarray experts,

i'm trying to run an microarray experiment with knock-out embryos. my desired genotype is Tg1/Tg1;Tg2/+ using Tg1/+;Tg2/+ and Tg1/+;+/+ as parents (
Tg1/Tg1 is non-viable so I can't use it as parents). Using this mating strategy, the chance I will get a embryo with a desired genotype is 1/8 in the whole litter, which means at best, I will get 1-2 embryos. I'm not sure if this is enough for a gene array experiment. so i'm considering to save the cells after sorting in RNAlater or progressing all the way to RNA extraction, and saving the RNA; then once enough cells or RNA has been accumulated then I will send it off for microarray. What would be the best way to accumulate cells/RNA for samples to be collected over time due to difficult-to-obtain tissues?

thanks so much for your response!!!

- minnie

#2 gogreen

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Posted 01 February 2011 - 01:53 AM

Hi Minnie, How much of RNA would you expect to obtain from one embryo in theory? Try to do one RNA isolation from an embryo and figure out what to expect in reality. I guess it's easier to do the RNA isolation with enough accumulated cells than trying to isolate small amounts of RNA over many trials! You can use RNAlater to store the embryos until you have enough to start with.

gg




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