Real time PCR profiling of 330 human micro-RNAs.
The small size of miRNAs makes profiling of all the 462 known human miRNAs difficult using single cell samples. Recently, we showed that judicious sequence partitioning between RT primers and second strand synthesis primers permitted multiplexed RT-PCR amplification of miRNA in very small samples to allow individual real time PCR quantification. Here, we show that zip coding the primers and TaqMan probes with sequences specific to each miRNA greatly improves reaction specificity, which permits the profiling of all miRNAs in a single multiplexed RT-PCR reaction.
Lao
Kaiqin
K
Applied Biosystems, Foster City, CA, USA. laokq@appliedbiosystems.com
Xu
Nan Lan
NL
Sun
Yongming Andrew
YA
Livak
Kenneth J
KJ
Straus
Neil A
NA
D001483:Base Sequence; D003199:Computer Systems; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D015347:RNA Probes / Q000235:genetics*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017423:Sequence Analysis, RNA / Q000379:methods*
D035683:MicroRNAs; D015347:RNA Probes