DyNAmo™ SYBR^® Green 2-Step qRT-PCR Kit and
DyNAmo™ Capillary SYBR^® Green 2-Step qRT-PCR Kit

For reliable RNA quantification

Description

Finnzymes' DyNAmo™ SYBR® Green 2-Step qRT-PCR Kit and DyNAmo™ Capillary SYBR® Green 2-Step qRT-PCR Kit enable reproducible detection and quantification of RNA with great sensitivity and specificity. The kits includes all the necessary reagents for cDNA synthesis and following SYBR Green qPCR. 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 version of a modified Thermus brockianus (Tbr) DNA polymerase. A non-specific DNA-binding domain has been fused to the polymerase, which lends physical stability to the polymerase-DNA complex. DyNAmo SYBR Green 2-step qRT-PCR Kit is compatible with all block-based real-time qPCR instruments and DyNAmo™ Capillary SYBR® Green 2-Step qRT-PCR Kit is engineered for capillary-based 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: uniquely engineered Tbr DNA polymerase.
• Convenient to use: 2x mixes and short protocols.

Broad dynamic range

Amplification and standard curves below (Figures 1a and 1b) show the wide dynamic range and excellent linearity of the DyNAmo SYBR Green 2-Step qRT-PCR Kits.

Figures 1a and 1b. Human HeLa cell total RNA amounts varying from 10 pg up to 1 ug were reverse transcribed using random priming. In the following qPCR 140 bp Calmodulin 1 fragment was amplified with cDNA specific primers. The cDNA synthesis reactions were done in 20 µl volumes, from which 1/14 part was transferred to qPCR reactions. Reactions were set up on the DNA Engine Opticon® system (Bio-Rad Laboratories).

Components

Both DyNAmo SYBR Green 2-Step qRT-PCR Kits are available in two sizes.

DyNAmo™ SYBR® Green 2-Step qRT-PCR Kit	F-430S	F-430L
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 (containing hot start version of modified Thermus brockianus DNA polymerase, SYBR Green I dye, optimized PCR buffer, MgCl2 and dNTP mix including dUTP) 50x ROX™ passive reference dye	40 qPCR reactions (50 µl each)	200 qPCR reactions (50 µl each)

DyNAmo™ Capillary SYBR® Green 2-Step qRT-PCR Kit	F-440S	F-440L
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 (containing hot start version of modified Thermus brockianus DNA polymerase, SYBR Green I dye, optimized PCR buffer, MgCl2 and dNTP mix including dUTP)	100 qPCR reactions (20 µl each)	500 qPCR reactions (20 µl each)

Storage and stability

Store all kit components at -20 °C. Both DyNAmo SYBR Green 2-Step qRT-PCR Kits are stable for six months from the date of packaging when stored and handled properly.

Ordering information

DyNAmo™ SYBR® Green 2-Step qRT-PCR Kit
F-430S	20 RT reactions (20 µl each) and 40 qPCR reactions (50 µl each)
F-430L	100 RT reactions (20 µl each) and 200 qPCR reactions (50 µl each)
DyNAmo™ Capillary SYBR® Green 2-Step qRT-PCR Kit
F-440S	20 RT reactions (20 µl each) and 100 qPCR reactions (20 µl each)
F-440L	100 RT reactions (20 µl each) and 500 qPCR reactions (20 µl each)