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The exponential amplification via reverse transcription polymerase chain reaction provides for a highly sensitive technique in which a very low copy number of RNA molecules can be detected. RT-PCR is widely used in the diagnosis of genetic diseases and, semiquantitatively, in the determination of the abundance of specific different RNA ...
A reverse transcriptase (RT) is an enzyme used to convert RNA genome to DNA, a process termed reverse transcription.Reverse transcriptases are used by viruses such as HIV and hepatitis B to replicate their genomes, by retrotransposon mobile genetic elements to proliferate within the host genome, and by eukaryotic cells to extend the telomeres at the ends of their linear chromosomes.
Nucleic acid amplification is a technique used to produce several copies of a specific segment of RNA/DNA. [3] Amplified RNA and DNA can be used for a variety of applications, such as genotyping, sequencing, and detection of bacteria or viruses. [4] There are two different types of amplification, non-isothermal and isothermal. [5]
This allows the virus sample to be grown into larger quantities and allows a larger number of tests to be run on them. This is particularly important for samples that contain new or rare viruses for which diagnostic tests are not yet developed. [citation needed] Many viruses can be grown in cell culture in the lab.
Total DNA and/or RNA are extracted from the samples and are prepared on a DNA or RNA library for sequencing. [9] These methods have been used to sequence the whole genome of Epstein–Barr virus (EBV) and HCV , however, contaminating host nucleic acids can affect the sensitivity to the target viral genome with the proportion of reads related to ...
In addition, human immunodeficiency virus, a single-stranded RNA virus, encodes an RNA-dependent DNA polymerase (reverse transcriptase) that uses complementarity to catalyze genome replication. The reverse transcriptase can switch between two parental RNA genomes by copy-choice recombination during replication.
There are also a number of RNA-dependent RNA polymerases that use RNA as their template for synthesis of a new strand of RNA. For instance, a number of RNA viruses (such as poliovirus) use this type of enzyme to replicate their genetic material. [58] Also, RNA-dependent RNA polymerase is part of the RNA interference pathway in many organisms. [59]
Portions of the enzyme were made transparent so as to make the path of RNA and DNA more clear. The magnesium ion (yellow) is located at the enzyme active site. The 17-bp transcriptional complex has an 8-bp DNA-RNA hybrid, that is, 8 base-pairs involve the RNA transcript bound to the DNA template strand. [17]
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