RTPCR Full Form
RTPCR Full Form-What Is RTPCR And How Does It Work? RTPCR is a laboratory technique combining reverse transcription of RNA into DNA and amplification of specific DNA targets using polymerase chain reaction.
|Reverse transcription polymerase chain reaction|
FAQs About RTPCR:
Reverse transcription polymerase chain reaction or RTPCR is a laboratory technique combining reverse transcription of RNA into DNA and amplification of specific DNA targets using polymerase chain reaction. Reverse transcription is the process of creating RNA, which is then transcribed back into DNA and amplified. RTPCR is one of the most commonly used laboratory techniques and has been around since the 1950s. One application of this technique is to detect bacteria by testing for their specific RNA molecules. Another common application of RTPCR is to determine a microbe’s antibiotic resistance.
Upon injection of the desired DNA sample into the sample chamber, RTPCR is used to perform the following steps: All of the following reactions proceed at an exponential rate: A DNA template containing known gene sequences, the targets of the PCR reaction, is amplified using the primers that have been specifically designed for this task. Each primer contains a sequence that contains the DNA sequence to be amplified and a sequence that recognizes a tag for the corresponding target gene. The sequencing template is converted into DNA following this PCR reaction. Following amplification of the desired sequence, the transformed DNA is hybridized with specific PCR antibody. The antibody binds the target gene, and the amplified DNA (or template DNA) is then isolated from the PCR reaction.
In the biological test field, RTPCR can be used to profile genetic sequences, and investigate the effects of genotype on the phenotype (the observable characteristics of a biological system). RTPCR tests are most commonly used for genomic sequencing for medical diagnostic applications, epigenetic studies in a variety of diseases, and cancer studies. Different types of RTPCR applications exist, and each have their particular advantages and disadvantages. These include De Novo amplification of short sequences: This type of RTPCR works very well for amplifying short, diverse regions of DNA to lower copy numbers. De novo amplification of long sequences: Unlike short sequences, long sequences are difficult to amplify in the laboratory, requiring expensive antibody reagents and controls.
Though this technique is valuable in cancer diagnosis and diagnosis of other diseases, its use is limited as it can only target specific sequences of the genome. Moreover, there is also uncertainty regarding the length of a target sequence that RTPCR can detect. The European Bioinformatics Institute states that RTPCR is limited to 3kb targets, this is a function of the fact that it deals with single-stranded DNA and it has difficulty detecting complex long sequences such as oncogenes and tumor suppressors. Targets have to be close to each other and in the same order to be detected by RTPCR. When RTPCR does not detect target it uses an alternative protocol.
RNA-based testing has become an integral part of the clinical diagnostics market, and the global market is expected to grow at a steady rate. With the advancements in molecular diagnostics, the global market for RNA-based diagnostics is expected to grow at a steady pace in the near future.