TRANSCRIPTME is a modified, recombinant form of the Reverse Transcriptase from Moloney Murine Leukemia Virus (M-MuLV) purified from Escherichia coli. TRANSCRIPTME Reverse Transcriptase synthesizes a complementary DNA strand in the presence of a primer using either RNA (cDNA synthesis) or single-stranded DNA (ssDNA) as a template.
The TRANSCRIPTME RNA Kit has been formulated to provide high yields of full-length cDNA product and to increase sensitivity in RT-qPCR. Starting material can range from 10 pg to 5 μg of total RNA.; optimal reaction temp. 50°C; contains Enzyme Mix (M-MuLV reverse transcriptase and RNase inhibitor); 2x Master Mix (oligodT primers, random hexamers, dNTPs, MgCl2) and RNase H in a separate tube (optional step)
RNase H is a 18.9 kDa recombinant endoribonuclease purified from an Escherichia coli strain. The enzyme hydrolyses specifically the phosphodiester bonds of RNA hybridized to DNA and produces 5´ phosphate-terminated oligoribonucleotides and single-stranded DNA. The enzymes does not degrade single and double-stranded DNA or unhybridized RNA. It is a key enzyme in the removal of mRNA after first-strand cDNA synthesis. Treating cDNA with RNase H prior to PCR can improve sensitivity as RNA bonded to the cDNA template may prevent binding of the amplification primers in a PCR reaction. RNase treatment is often necessary when amplifying longer, full-length cDNA targets. In addition, this enzymes is useful for the removal of poly(A) tails on mRNAs after hybridization with oligo(dT) and also for the site-specific enzymatic cleavage of RNA.