Properties
Form: solution
Quality level: 100
Specific activity
- >40 units/μg of protein
- >40 units/mg
Packaging: 500u pack
Manufacturer/trade name: Roche
Sent in: dry ice
Storage temperature: −20°C
General description
M-MuLV reverse transcriptase (from Moloney murine leukaemia virus) is produced as a cloned enzyme resulting from the fusion of the Escherichia coli type gene with the core region of the M-MuLV pol gene. The enzyme is expressed in E. coli HB101 pB6B 15.23. The enzyme is an RNA-dependent DNA polymerase that uses single-stranded RNA or DNA as a template in the presence of a primer to synthesize a complementary DNA strand.
Compared with AMV (avian myeloblastosis virus) reverse transcriptase, M-MuLV reverse transcriptase lacks endonuclease activity and has much lower RNase H activity. Full copies of large mRNA are obtained. The M-MuLV reverse transcriptase contains an N-terminal DNA polymerase domain, a connecting domain, and a C-terminal RNase H domain. Genome sequence-specific degradation by M-MuLV reverse transcriptase is considered an efficient method for targeting drugs specific for retroviral H.
M-MuLV reverse transcriptase is produced as a cloned enzyme resulting from the fusion of the E. coli type gene with the core region of the M-MuLV pol gene. The enzyme is expressed in E. coli HB101 pB6B 15.23. The enzyme is an RNA-dependent DNA polymerase that uses single-stranded RNA or DNA as a template in the presence of a primer to synthesize a complementary DNA strand. Compared to AMV reverse transcriptase, M-MuLV reverse transcriptase lacks endonuclease activity and has much lower RNase H activity. Full copies of large mRNA are obtained.
Specificity
M-MuLV reverse transcriptase is produced as a cloned enzyme resulting from the fusion of the E. coli type gene with the core region of the M-MuLV pol gene. The enzyme is an RNA-dependent DNA polymerase that uses single-stranded RNA or DNA as a template in the presence of a primer to synthesize a complementary DNA strand.
Request
Reverse transcriptase M-MuLV (from Moloney murine leukaemia virus) can be used to:
- In vitro synthesis of cDNA transcripts of specific RNA sequences
- Preparation of cDNA libraries
- Dideoxy sequencing reactions in place of the Klenow enzyme. M-MuLV reverse transcriptase will often be synthesized through GC-rich regions where the Klenow enzyme is hampered
- Transcription of a simple and abundant RNA target.
The cDNA products are used for the structure, organization, and expression analysis of prokaryotic and eukaryotic genes. Comparison between cDNA and genomic DNA sequences elucidates intervening sequences, splicing events, and genomic rearrangements in eukaryotic genes. M-MuLV reverse transcriptase has been used for 3′ and 5′ RACE (rapid amplification of cDNA ends). It has also been used for the synthesis of first-strand cDNA for use in downstream amplification reactions (RT-PCR)
Features and Benefits
Sturdiness:
- Obtain cDNA transcripts up to 10 kb
- No endonuclease activity
- Lower RNase H activity than AMV (avian myeloblastosis virus)
- Efficiently transcribes total RNA, mRNA, viral RNA, and RNA rich in secondary structures
