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 1a
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.

	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. 5'-amplicon 3'-amplicon
	Company I RNase H- minus MMLV RT
	Company P MMLV RT

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Protocols for: MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit 

Due to the constant updating of the protocols by the manufacturer we have provided a direct link to Lucigen's product page, where the latest protocol is available.

Please note this will open a new page or window on your computer.

MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit Protocol

(catalogue number  MM070150) 

MMLV High Performance Reverse Transcriptase Protocol

(catalogue number RT80125K)

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References:

1. Catalano, P. N., et al. (2010) Lack of functional GABAB receptors alters GnRH physiology and sexual dimorphic expression of GnRH and GAD-67 in the brain, Am J Physiol Endocrinol Metab 298 , E683-696.
2. Chen, T.-L., et al. (2010) Contribution of a Plasmid-Borne blaOXA-58 Gene with Its Hybrid Promoter Provided by IS1006 and an ISAba3-Like Element to {beta}-Lactam Resistance in Acinetobacter Genomic Species 13TU, Antimicrob. Agents Chemother. 54 , 3107-3112.
3. Tan, D. & Walker, A. M. (2010) Short form 1b human prolactin receptor down-regulates expression of the long form, J. Mol. Endocrinol. 44 , 187-194.
4. Tian, Y., et al. (2010) MicroRNA-10b Promotes Migration and Invasion through KLF4 in Human Esophageal Cancer Cell Lines, J. Biol. Chem. 285 , 7986-7994.
5. Wu, Z.-B., et al. (2010) Biological and Molecular Characterization of Apple chlorotic leaf spot virus Causing Chlorotic Leaf Spot on Pear (Pyrus pyrifolia) in Taiwan, HortScience 45 , 1073-1078.
6. Xin, Q.-Q., et al. (2010) Apoptosis contributes to testicular toxicity induced by two isomers of bromopropanes, Toxicology and Industrial Health 26 , 513-524.
7. Chen, G.-Y., et al. (2009) Baculovirus Transduction of Mesenchymal Stem Cells Triggers the Toll-Like Receptor 3 Pathway, J. Virol. 83 , 10548-10556.

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Applications

• First-strand cDNA synthesis for subsequent PCR or real-time PCR.
• RT-PCR validation of gene expression data obtained from microarray experiments.
• RT-PCR validation and quantification of gene silencing by RNA interference

Benefits

• Synthesize full-length cDNA from RNA templates longer than 15 kb.
• MMLV HP RT demonstrates significantly higher activity than competitive reverse transcriptase enzymes.
• Reaction Buffer, optimized for producing full-length cDNA, is included with both the MMLV HP RT and the MMLV Reverse Transcriptase 1st-Strand cDNA Synthesis Kit.
• The kit includes both an oligo(dT) and random nonamer primers.
• First-strand cDNA can be made from picogram amounts of input total RNA.
• The kit includes a potent RNase Inhibitor to protect the integrity of template RNA.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200