The MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit is optimized for generating full-length first-strand cDNA from total cellular RNA preparations or purified polyadenylated RNA.
MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit includes Epicentre's high activity native MMLV reverse transcriptase and an optimized, proprietary Reaction Buffer. A kit reaction is capable of synthesizing 1st-strand cDNA from RNA templates longer than 15 kb.
|Figure 1. The MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit produces full-length cDNA from mRNA longer than 15 kb. Total RNA isolated from HeLa cells was reverse transcribed and the cDNA was amplified as described in the text. A. Detection of the 1.3 kb PCR amplicon from near the 5' end of the mRNA demonstrates full-length reverse transcription of HERC1 mRNA. B. Agarose gel analysis of the PCR reaction shows the 1.3 kb amplicon from the 5'-end of the mRNA. Lane M, 100 bp DNA ladder; Lane 1, no reverse transcriptase control reaction; Lane 2, PCR product from cDNA synthesized by Epicentre's MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit.
Table 1. 3'/5' ratio analysis of cDNA produced by different reverse transcriptase enzymes. Total cellular RNA from HeLa was converted to cDNA using the three reverse transcriptase enzymes indicated in the table. A 3'/5' ratio equal to 1.0 means that equal amounts of PCR products are obtained from both the 3'- and 5'-end of the cDNA and therefore is a good indication that the reverse transcriptase has produced a full-length cDNA copy of the mRNA.
Target Transcript (Size)
EPICENTRE MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit
Competitior I (RNaseH- mutant of MMLV RT)
Competitor P (MMLV RT)
|3'/5' ratio ACTB (1792 b)
|3'/5' ratio GUSB (2162 b)
|3'/5' ratio TFRC (5010 b)
|Epicentre MMLV 1st-Strand cDNA Synthesis Kit
|Figure 2. cDNA produced by Epicentre's MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit yields a significantly improved 3'/5' ratio than competitive reverse transcriptases. The approximately 2160-base HeLa β-glucuronidase mRNA (GUSB) was reverse transcribed into cDNA using Epicentre's MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit and two competitive reverse transcriptase enzymes. PCR primer pairs to the 3'-end and 5'-end of GUSB cDNA were synthesized and qPCR (SYBR® Green I dye detection) was performed using each primer pair and the GUSB cDNAs as templates. The 3'/5' ratio was calculated for each as described in the text. A. PCR amplicons from the 3' end and 5' end of the GUSB cDNA B. qPCR quantification graphs for detecting the 3' amplicon and 5' amplicon of GUSB cDNA produced by Epicentre's MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit and two other commercially available reverse transcriptases.
|Company I RNase H- minus MMLV RT
|Company P MMLV RT
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