Properties

Form: solution

Quality level: 100

Specific activity

> 40 units / μg protein
> 40 U / mg

Packaging: 500 U pack

Sent in: dry ice

Storage temperature: −20 ° C

General description

The 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 central 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 RNA or single-stranded DNA as a template in the presence of a primer to synthesize a complementary DNA strand.

Compared to AMV (Avian Myeloblastosis Virus) reverse transcriptase, M-MuLV reverse transcriptase lacks endonuclease activity and has much lower RNase H activity. Complete 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. Sequence-specific genome degradation by M-MuLV reverse transcriptase is considered an effective method of targeting drugs specific to retroviral H RNases.

The M-MuLV reverse transcriptase is produced as a cloned enzyme resulting from the fusion of the E. coli type gene with the central 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 RNA or single-stranded 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. Complete copies of large mRNA are obtained.

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Specificity

M-MuLV reverse transcriptase is produced as a cloned enzyme resulting from the fusion of the E. coli type gene with the central region of the M-MuLV pol gene. The enzyme is an RNA-dependent DNA polymerase that uses RNA or single-stranded DNA as a template in the presence of a primer to synthesize a complementary DNA strand.

Application

M-MuLV reverse transcriptase (from Moloney murine leukemia virus) can be used to:

• In vitro synthesis of cDNA transcripts of specific RNA sequences
• Preparation of cDNA libraries
• Dideoxy sequencing reactions instead of the Klenow enzyme. M-MuLV reverse transcriptase is often
• synthesized through GC-rich regions where the Klenow enzyme is hampered
• Simple and abundant RNA transcription.

The cDNA products are used for the analysis of the structure, organization and expression of prokaryotic and eukaryotic genes. Comparison between cDNA and genomic DNA sequences clarifies intermediate sequences, splicing events, and genomic rearrangements in eukaryotic genes. The 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 subsequent amplification reactions (RT-PCR).

Features and Benefits

Sturdiness:

• Get 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

Definition of unit

1. 1 unit is the enzymatic activity that incorporates 1.0 nmol of [3H] -TMP into an acid-insoluble product in 10 minutes at +37 ° C with poly (A) + x (DT) 15 as substrate.
2. Volume activity:> 20 U / μl. See certificate of analysis

Preparation note

• Storage conditions (working solution): – 70 ° C. The diluted solution can be stored in aliquots at -70 ° C.
• Storage buffer: 50 mM Tris-HCl, pH 8.4 (+ 4 ° C), 10 mM dithioerythritol, 100 mM NaCl, polidocanol, 0.05% (v / v), 1 mM EDTA, 50% glycerol, (v / v).

Other notes

For life science research only. It should not be used in diagnostic procedures.

Reaction requirements

The M-MuLV reverse transcriptase reaction requires a primer.