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Pooling doesn't work


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#1 wntiong

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Posted 23 August 2009 - 07:14 PM

Hi, I want to increase amount of my rna so that i can get a total of 10ug for my microarray run. However, each aliquot that i have only has higher amount of 6-8ug. I thought pooling could help, however after i pooled my 4 aliquots (which each yield abt 4-5ug), the result of pooled sample is only 1.8ug! I lost much of rna during precipitation. Any alternative despite of IVT amplification kit can help me to get enough rna for my microarray run? [my sample is enough, is just that i dunno how to make up to 10 ug]. =)

Thanks

#2 stylothecancer

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Posted 24 September 2009 - 12:24 AM

Hi, I want to increase amount of my rna so that i can get a total of 10ug for my microarray run. However, each aliquot that i have only has higher amount of 6-8ug. I thought pooling could help, however after i pooled my 4 aliquots (which each yield abt 4-5ug), the result of pooled sample is only 1.8ug! I lost much of rna during precipitation. Any alternative despite of IVT amplification kit can help me to get enough rna for my microarray run? [my sample is enough, is just that i dunno how to make up to 10 ug]. =)

Thanks



Hi,

Have you figure out how to solve your problem???

#3 wntiong

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Posted 24 September 2009 - 12:33 AM

Hi, I want to increase amount of my rna so that i can get a total of 10ug for my microarray run. However, each aliquot that i have only has higher amount of 6-8ug. I thought pooling could help, however after i pooled my 4 aliquots (which each yield abt 4-5ug), the result of pooled sample is only 1.8ug! I lost much of rna during precipitation. Any alternative despite of IVT amplification kit can help me to get enough rna for my microarray run? [my sample is enough, is just that i dunno how to make up to 10 ug]. =)

Thanks



Hi,

Have you figure out how to solve your problem???



Hi, I pooled the pellets from several aliquots during rna extraction and it worked, finally. but need to perform several times of etoh washing in order to get a pure rna sample

#4 stylothecancer

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Posted 24 September 2009 - 02:06 PM

Great to hear that. :wacko:

I am sure you are aware of by pooling of RNA samples for microarray, you are susceptible to some dilution effect to your microarray data later if you are not proper with you experimental design. :)




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