DyNAmo™ Probe 2-Step qRT-PCR Kit

For reliable RNA quantification

Description

Finnzymes' DyNAmo™ Probe 2-Step qRT-PCR Kit enables reproducible detection and quantification of RNA with great sensitivity and specificity. The kit includes all the necessary reagents for cDNA synthesis and following qPCR. Only template RNA, PCR primers and probe need to be added by the user. The reverse transcriptase in the kits is M-MuLV RNase H+, which provides higher sensitivity to qPCR than RNase H- reverse transcriptases. The RNase H activity in the RT enzyme facilitates annealing of PCR primers to the cDNA by degrading the RNA template from the RNA-cDNA hybrid before the PCR step. The performance of the qPCR step is based on a hot start Thermus brockianus (Tbr) DNA polymerase. DyNAmo Probe 2-step qRT-PCR Kit is compatible with all block-based and capillary-based real-time qPCR instruments.

Advantages

• Broad dynamic range: up to 1 µg of RNA.
• Reliable results with different RNA sources: mRNA, total RNA, viral RNA and in vitro transcribed RNA.
• Flexible: two-step system, all priming options.
• Optimized 2x mixes for both cDNA synthesis and qPCR.
• Specific and sensitive: efficient Tbr DNA polymerase.
• Convenient to use: 2x mixes and short protocols.
  

Reliability, specificity and sensitivity

Human HeLa cell total RNA amounts varying from 10 pg to 1µg were reverse transcribed using oligo(dT) priming. The cDNA synthesis reactions were done in 20 µl volumes, from which 1/14 was transferred to qPCR reactions. A Calmodulin 1 fragment was amplified with cDNA specific primers and detected with an LNA probe (ProbeLibrary™, Exiqon A/S). Reactions were set up on DNA Engine Opticon® system (Bio-Rad, Inc.).

Components

DyNAmo™ Probe 2-Step qRT-PCR Kit	F-460S	F-460L
cDNA synthesis: M-MuLV RNase H+ reverse transcriptase (includes RNase inhibitor) 2x RT buffer (includes dNTP mix and MgCl2 Random hexamers (300 ng/µl) oligo(dT)15 primer (100 ng/µl)	20 RT reactions (20 µl each)	100 RT reactions (20 µl each)
qPCR: 2x master mix (hot start Tbr DNA polymerase, optimized PCR buffer, MgCl2, dNTP mix including dUTP) 50x ROX™ passive reference dye	40 qPCR reactions (50 µl each)	200 qPCR reactions (50 µl each)

Storage

Store all kit components at -20°C.

Ordering information

DyNAmo™Probe 2-Step qRT-PCR Kit
F-460S	20 RT reactions (20 µl each) and 40 qPCR reactions (50 µl each)
F-460L	100 RT reactions (20 µl each) and 200 qPCR reactions (50 µl each)