FastGene Scriptase II cDNA Kit

FastGene® Scriptase II cDNA Kit (100 rxns) contains FastGene Scriptase II, enzyme buffer, dNTPs, Oligo dTs, random hexamers and RNase inhibitor

Engineered enzyme

Engineered reverse transcriptases allowed the synthesis of cDNA from very low amounts of RNA. Mutations are inserted into the RNase H domain of the MuLV‘s reverse transcriptase. Therefore, by not degrading the RNA during the first-strand synthesis, a higher yield of full-length cDNA is obtained.

Additionally, a higher thermal stability increases the robustness of the enzyme. The FastGene® Scriptase II is exactly one of those engineered enzymes. With its mutation in the RNase H domain and higher thermal stability, it is the optimal choice for more complex applications, such as RT-qPCR and NGS.

Applications

• Quantification of Gene Expression
• RT-qPCR
• Next Generation Sequencing
• Low RNA concentration
• Difficult templates

Lower RNase H activity for longer cDNA

The FastGene® Scriptase II has a modified RNase H domain. The RNA is therefore not degraded and serves as a template for longer cDNAs, resulting in fragment size of up to 12 kBp.
Engineered enzymes – optimized for qPCR

The FastGene® Scriptase II delivers superior cDNA templates for downstream applications, e.g. qPCR and NGS. The resulting full-length cDNA gives a complete picture of the gene and is able to show modification, e.g. splicing variants.
Comparison of different PCR results

Multiplex PCR

Fig. 1: Comparison of multiplex PCR using cDNA produced by Competitor I‘s SS-II enzyme and FastGene® Scriptase II at 42 °C and 50 °C.

qPCR

Fig. 2: Comparison of qPCR results using primers for GAPDH and cDNA produced by using different RNA starting concentration by Competitor I‘s SS-II enzyme   and FastGene® Scriptase II at 42 °C.

Fig. 3: Comparison of qPCR results using primers for YWHAZ and cDNA produced by using different RNA starting concentration by Competitor I‘s SS-II enzyme and FastGene® Scriptase II at 42 °C.

Kit components:

• FastGene®Scriptase II (200 U/μl)
• 5x FastGene® RT buffer
• DTT (0.1 M)
• dNTP Mixture (2 mM each)
• Sterile water (RNase free)
• RNase Inhibitor (40 U/μl)
• Oligo dT Primer (80 μM)
• Random hexamer (100 μM)