cy3 and cy5-dCTP labelling

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bsengez
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cy3 and cy5-dCTP labelling

Post by bsengez » Thu Jul 22, 2010 12:54 pm

I purified total RNA from two different cell lines stably expressing an empty vector (as a control) and vector having two inserts. Now, I want to perform microarray. We have Cy3-dCTP and CY5-dCTP. Some protocols want you to purify mRNA and reverse transcribe it to cDNA, the others use total RNA to make cDNA. Which one is more suitable for these dyes? Is anyone using these dyes for cDNA or RNA labelling? I need a good protocol for it.
Thanks in advance..
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JackBean
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Post by JackBean » Thu Jul 22, 2010 6:34 pm

IMHO that doesn't matter. Just in case of total RNA you must use higher amount of RNA, as the mRNA is only small fraction
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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bsengez
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Post by bsengez » Thu Jul 22, 2010 6:42 pm

How much RNA do I need to start with? Do you have any protocols for cDNA labelling?
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