TRANSCRIPTME RNA Kit - cDNA synthesis kit, 20 reactions supplied by BLIRT is a system having all the necessary components for synthesizing first-strand cDNA, apart from the template total RNA or mRNA. The synthesized single-stranded cDNA is suitable for real-time quantitative RT-PCR applications. The TRANSCRIPTME RNA Kit gives a high yield of full-length cDNA product which increases sensitivity in RT-qPCR. Starting material can range from 10 pg to 5 g of total RNA. The kit contains TRANSCRIPTME Enzyme Mix, with a recombinant thermostable reverse transcriptase M-MuLV without RNase H activity and the RIBOPROTECT RNase Inhibitor along with the optimal reaction buffer with a combination of random hexamers and oligo(dT)18 primers, MgCl2 and dNTPs for increased sensitivity. TRANSCRIPTME Reverse Transcriptase is highly thermally stable due to which the reaction can be set up at higher temperatures (optimum activity at 50C). It enhances the effectiveness and specificity of those transcribed RNA regions which are rich in GC pairs and/or contain secondary structures. The enzyme is devoid of 35 exonuclease or RNase H activitywhich improves synthesis of a full-length cDNA, even from long mRNA templates, using random priming. The enzyme gives high yields of first strand cDNA up to 10 kb long. Using the recombinant RNase inhibitor RIBOPROTECT prevents the RNA template from getting degraded. The enzyme only hydrolyze the phosphodiester bonds of RNA when hybridized to DNA. The RNase H does not degrade either single and double stranded DNA or unhybridized RNA. This optional step can improve the sensitivity of subsequent RT-qPCR reaction since the PCR primers will bind more easily to the cDNA. Using of the RNase H is recommended only when it contributes to a full-length cDNA synthesis and increased yields of first-strand cDNA.

Applications include:

Full-length cDNA template synthesis for RT-qPCR and two-step RT-PCR assays

cDNA synthesis for molecular cloning

cDNA library construction

RNA analysis

Product Specifications:

Product name - TRANSCRIPTME RNA Kit - cDNA synthesis kit, 20 reactions

Catalog No - RT31-020

Brand: BLIRT

Pack size: 20 reactions

Product Features:

High yields of full-length cDNA products (up to 10 kb)

Formulated to increase sensitivity in RT-qPCR and RT-PCR assays

Fewer pipetting steps minimized contamination risk

Simultaneous cDNA synthesis with mRNA and rRNA templates

Starting material: 10 pg 5 g of total RNA or 10 pg 500 ng of mRNA

Optimal reaction temperature: 50C

The reverse transcriptase shows no RNase H or 35 exonuclease activity, has supreme thermostability and is suitable for difficult RNA templates

RNase inhibitor protects the RNA template from degradation

Additional RNase H treatment may increase the sensitivity of the RT-qPCR reaction.