PrimeScriptTM Reverse Transcriptase

Description

PrimeScript Reverse Transcriptase is a modified MMLV Moloney Murine Leukemia Virus RTase. This enzyme has extremely high extension capability and can synthesize long first-strand cDNA efficiently. Even for difficult templates, including RNAs with complex secondary structure, it is possible to synthesize first-strand cDNA at the normal reverse transcription temperature 42C with this enzyme. It is not necessary to perform the RT reaction at higher temperatures a condition that may cause RNA degradation . This enzyme is suitable for preparation of long cDNAs, for construction of cDNA libraries that include full-length cDNA, etc. Cat. # 2680B contains 4 of Cat. # 2680A. Please refer to Cat. # 2680A for complete product documentation and resources. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Strand displacement and elongation activity of PrimeScript Reverse Transcriptase in comparison to Company A RTase.Top panel: diagram of cDNA synthesis reactions performed using either an oligo dT primer resulting in a 6.4-kb product or a gene-specific primer yielding a 4.4-kb product . Primer extension reactions were performed at 50C. Bottom panel: Primer extension reactions were analyzed on an alkaline denaturing gel. PrimeScript Reverse Transcriptase provided higher yield of both the 6.4-kb and the 4.4-kb product, demonstrating superior strand displacement capability. Back cDNA synthesis with low background, excellent extension, and high yield at 42C by PrimeScript Reverse Transcriptase in comparison to six other commercially available RTases.RNA ladder 1 kb, 2 kb, 4.4 kb, 6.4 kb, 8.4 kb, 10 kb, and 12 kb was used as template for first-strand cDNA synthesis reactions. Reactions were performed at 42C according to the recommendations of each manufacturer. Equivalent reaction volumes were analyzed using alkaline denaturing gel electrophoresis and products were detected with green intercalating dye and fluorescence image analysis. Reactions performed using PrimeScript Reverse Transcriptase showed the highest yield of full-length cDNA product and lowest background. Back Highly accurate reverse transcription with PrimeScript Reverse Transcriptase.First-strand cDNA synthesis reactions were performed using PrimeScript Reverse Transcriptase or other commercially available RTases. The template for all reactions was Human Placenta Total RNA Takara Bio . All reactions used an oligo dT primer and were performed according to the recommendations of each manufacturer. After synthesizing cDNA, PCR amplification was performed with highly accurate PrimeSTAR HS DNA Polymerase target gene: TF, amplification product: 500 bp . A control reaction was included in which PCR was performed no reverse transcription step . Amplified fragments were cloned and multiple clones from each reaction were sequenced. Error rate was defined as the number of errors per total bases sequenced for each reaction type 200,000 bases . With only 7 errors out of 201,297 bases 0.0035 % error rate , PrimeScript Reverse Transcriptase showed the highest accuracy of all RTases studied. Back