Reverse Transcriptase is an engineered version of M-MLV reverse transcriptase with deficient RNase H activity. The enzyme is purified to near homogeneity from E.coli containing the modified M-MLV RT gene.
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- Category: RT-PCR
What is reverse transcriptase?
Reverse transcriptase (RT), also known as RNA-dependent DNA polymerase, is a DNA polymerase that transcribes single-stranded RNA into DNA. Once RNA is reverse transcribed into single-stranded DNA in the first step, the enzyme is able to synthesize double-helix DNA.
Reverse transcriptase is an RNA-dependent DNA polymerase that was discovered in 1970 in many retroviruses including human immunodeficiency virus (HIV) and avian myeloblastosis virus (AMV). Reverse transcriptase catalyzes the conversion of RNA template molecules into DNA double helices and provides a very useful tool for molecular biology research.
Reverse transcriptase is commonly used to generate complementary DNA (cDNA) libraries from variously expressed mRNAs and, when used in conjunction with a polymerase chain reaction technique called RT-PCR, can also be used to quantify levels of mRNA synthesis.
The function of reverse transcriptase
Transcriptase (RT) is an enzyme used to generate complementary DNA (cDNA) from RNA templates, a process known as reverse transcription. Viruses such as HIV and Hepatitis B use reverse transcriptase to replicate their genomes, proliferate within the host genome by moving genetic elements through retrotransposons, and extend telomeres at the ends of their linear chromosomes by eukaryotic cells. Contrary to the commonly held view, In contrast, the process does not violate the flow of genetic information described by the classical central dogma, since the transfer of information from RNA to DNA is explicitly thought to be possible
Reverse transcriptase contains three enzymatic activities:
(1) RNA-dependent DNA polymerase activity,
(2) ribonuclease RNase H,
(3) DNA-dependent DNA polymerase activity.
First, an RNA-dependent DNA polymerase synthesizes a DNA strand complementary to the RNA template. RNase H then removes the RNA strand from the RNA-DNA hybrid duplex. The DNA-dependent DNA polymerase then completes double-stranded DNA synthesis. Unlike other DNA polymerases, reverse transcriptase lacks proofreading ability and therefore has a high error rate during DNA synthesis, with at most one error in 2000 base incorporations. The high error rate of viral reverse transcriptase provides a selective advantage for its survival in the host system.
Applications of reverse transcriptase:
Reverse transcriptase inhibitor
Because HIV uses reverse transcriptase to replicate its genetic material and create new viruses (part of the retroviral proliferation cycle), specific drugs are designed to disrupt this process, thereby inhibiting its growth. Collectively known as reverse transcriptase inhibitors, these drugs include the nucleoside and nucleotide analogs zidovudine (Retrovir), lamivudine (Epivir), and tenofovir (Viread), as well as non-nucleoside inhibitors, such as nevirapine (Viramune).
Reverse transcription polymerase chain reaction
Reverse transcriptase is often used in research to apply the polymerase chain reaction technique to RNA, a technique called reverse transcription polymerase chain reaction (RT-PCR). Classic PCR techniques can only be applied to DNA strands, but with the help of reverse transcriptase, RNA can be transcribed into DNA, making PCR analysis of RNA molecules possible. Reverse transcriptase is also used to create cDNA libraries from mRNA. The commercial availability of reverse transcriptase has greatly improved knowledge in the field of molecular biology, as well as other enzymes, which allow scientists to clone, sequence, and characterize RNA.
Supplier of Reverse Transcriptase:
GENETURE is a group company focus on the field of clinical diagnosis and life sciences,provide one stop solution of Nucleic Acid Extraction and Analysis,including solution of COVID-19. Main products including: Nucleic acid extraction or purification kit,Automatic nucleic acid extractor, PCR system, PCR kit, Magnetic beads, and lab consumables of 96 well deep plate,Magnetic rod comb,PCR tube,PCR plate,Pipette tips,centrifuge tubes.etc.
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Product Detail Contents:
|RT||10000 U||5×10000 U|
|5×ES RT Buffer||200 μl||5×200 μl|
|Anchored Oligo(dT)18 Primer (0.5 μg/μl)||50 μl||250 μl|
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