Efficient, Sensitive and Accurate

Highly efficient poly(A) tailing and reverse transcription in a single reaction tube provides uniform cDNA synthesis of microRNAs. The optimized reaction conditions and buffer components maximize cDNA yield using anywhere from several micrograms to picograms of input total RNA. The universal 3' tag sequence incorporated during reverse transcription enables the ease of scalable and accurate microRNA expression analysis by qPCR. Profile 1000 different microRNAs from a single reverse transcription reaction.

 

Sensitive

Start with total, Trizol-extracted RNA or fractionated small RNA samples. Achieve measurements from several micrograms to picograms of input RNA with excellent accuracy. Detect microRNA expression changes in a dynamic range of at least 6 log-fold differences.

 

Accurate

First strand cDNAs were synthesized using 500 ng total RNA from 18 different human tissues: adipose, bladder, brain, cervix, colon, esophagus, heart, kidney, liver, lung, ovary, placenta, prostate, skeletal muscle, small intestine, spleen, testes, and thymus.
The QuantiMir cDNAs from the 18 human tissues were balanced to yield equal Ct values for the U6 snRNA normalizing transcript (Below in Green bars). Real-time PCR results demonstrated this snRNA is uniformly expressed across the 18 tissues examined.
Assays specific for microRNA miR-1 demonstrate specific Heart and Skeletal expression (Below in Red bars) and assays for microRNA miR-122 clearly show specific Liver expression (Below in Blue bars).

 

Versatile

 

Kit Components

 

The QuantiMir RT kit also includes three reference qPCR assays for Human U6, Mouse U6 and Human/Mouse miR-16.  More information for the endogenous reference qPCR controls is located in the microRNA resources section.