Ultra Pure High-Temp Reverse Transcriptase

M-MLV (Moloney Murine Leukemia Virus) Reverse Transcriptase DNA polymerase uses DNA, single-stranded RNA, or an RNA:DNA hybrid (using a primer) to synthesize a complementary DNA strand. Wild-type and traditional Moloney Murine Leukemia Virus Reverse Transcriptases (MMLV-RT) are restricted to lower temperatures (up to 50℃) during cDNA synthesis. For many reverse transcription reactions, temperatures higher than 50℃ are beneficial. Higher temperatures reduce the chances of inhibition due to RNA secondary structures, while at the same time increasing primer binding specificity. These benefits lead to an increase in desired cDNA yield from the reverse transcription reaction. Ultra pure High-temp Reverse Transcriptase maintains its performance at high temperatures and outcompetes wild-type MMLV-RTs.Ultra pure High-temp Reverse Transcriptase is manufactured under stringent conditions to ensure that it is free of detectable levels of DNA, DNase, RNase, and Nickase to reduce the risk of false positive and false negative results in molecular diagnostic assays. Ultra pure High-temp Reverse Transcriptase should be used with other ultra pure DNA-free reagents, such as Ultra pure 10x Reverse Transcriptase Buffer and Ultra pure DNA-free Water, to minimize false positive and negative results and improved cDNA synthesis.Ultra pure High-temp Reverse Transcriptase was designed for use in highly-regulated industries in applications that require a high level of performance, lot-to-lot consistency, and high sensitivity without false results from reagent-borne contamination while meeting industry-driven critical regulations. To meet this requirements we have increased the quality level of our products. The M-Clarity Program defines product quality. It provides transparency so that you can choose, with confidence, suitable products that meet your needs with respect to quality and regulatory compliance, transparency, change notification, and documentation.

Unit Definition-One unit incorporates 0.1 nmol of TTP into double stranded product in 10 min at 37°C using poly(A):oligo dT as a template:primer evaluated with a fluorescent assay.

ApplicationsUltra pure High-temp Reverse Transcriptase can be used for:

• Reverse transcriptionFirst strand cDNA synthesis (up to 7 kb)
• Preparation of cDNA libraries
• One- or two-step RT-PCR and RT-qPCR
• Rapid Amplification of cDNA Ends (RACE)
• RNA-Seq
• For further manufacture

Ultra pure 10x Reverse Transcriptase Buffer is provided in larger SKU sizes for ease of use in high throughput RT-PCR applications and for further manufacture. Ultra pure 10x Reverse Transcriptase Buffer should be used with other ultra pure DNA-free reagents, such as Ultra pure High-temp Reverse Transcriptase and Ultra pure DNA-free Water, to minimize false positive and negative results.

Quality testing ensures the absence of detectable levels of contaminating DNA (≤ 1 copy of prokaryotic gDNA, ≤ 0.1 copy of eukaryotic gDNA, and ≤ 10 copies of plasmid DNA per 100 units of enzyme) as detected by qPCR using universal primers against 16S capable of detecting > 500,000 prokayotes, 18S capable of detecting > 110,000 eukaryotes, and Ori capable of detecting > 25,000 plasmids, respectively.