Recently, I do the microRNA expression profiling for two samples. One sample is from huh7
transfected with plasmid expressing virus protein, another is from huh7 transfected with
invalid plasmid as mock. I used the ABI Taqman low density microRNA array (TLDA) to screen
the two RNA samples and got the microRNA profile. Can I run TLDA once, then select some interesting
microRNAs to validate using Taqman single-well real-time qPCR ? Do I need run TLDA multireplicate ?
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microRNA profiling using ABI taqman low density microRNA array (TLDA)
Started by yueh76, Jun 01 2010 07:55 PM
1 reply to this topic
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Posted 07 June 2010 - 02:56 AM
yueh76, on Jun 2 2010, 05:55 AM, said:
Recently, I do the microRNA expression profiling for two samples. One sample is from huh7
transfected with plasmid expressing virus protein, another is from huh7 transfected with
invalid plasmid as mock. I used the ABI Taqman low density microRNA array (TLDA) to screen
the two RNA samples and got the microRNA profile. Can I run TLDA once, then select some interesting
microRNAs to validate using Taqman single-well real-time qPCR ? Do I need run TLDA multireplicate ?
transfected with plasmid expressing virus protein, another is from huh7 transfected with
invalid plasmid as mock. I used the ABI Taqman low density microRNA array (TLDA) to screen
the two RNA samples and got the microRNA profile. Can I run TLDA once, then select some interesting
microRNAs to validate using Taqman single-well real-time qPCR ? Do I need run TLDA multireplicate ?
You'll want to be VERY careful whan using TLDAs. false positive are not rare so maybe it is worth doing some replicates. since you're using cells RNA concentration and amount shoud not be a problem so my advice is to skip preamplification if u can. if not be sure to validate your results. Single Taqman assays are ok for validation, just be sure to do at least duplicates in your qPCR plate!
bye
Fiz
