Premium
Real time PCR profiling of 330 human micro‐RNAs
Author(s) -
Lao Kaiqin,
Xu Nan Lan,
Sun Yongming Andrew,
Livak Kenneth J.,
Straus Neil A.
Publication year - 2007
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200600119
Subject(s) - taqman , computational biology , microrna , biology , real time polymerase chain reaction , profiling (computer programming) , gene expression profiling , microbiology and biotechnology , genetics , gene , gene expression , computer science , operating system
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.