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This page is about Vectorette PCR. For PCR, see Polymerase chain reaction. For other uses, see PCR (disambiguation).

Steps involved in PCR

The polymerase chain reaction (PCR) was created during the 1980s. Since then, multiple variants of PCR have been created. Vectorette PCR is one of the variants that have been derived from PCR and was designed in 1988 and also patented. Vectorette PCR focuses on amplifying a specific sequence obtained from an internal sequence that is originally known until the fragment end. Multiple researches have taken this method as an opportunity to conduct experiments in order to uncover the potential uses that can be derived from Vectorette PCR.

Introduction

Vectorette PCR is similar to PCR; however, it is capable of obtaining the sequence desired for amplification from an already known primer site. While PCR needs information of already known sequences at both ends, Vectorette PCR only requires previous knowledge of one.This means that is able to apply the method of PCR which needs sequence information from both ends to fragments of DNA that contain the information of the sequence at only one end and not the other. In order to achieve this, there are specific steps that this method must first go through. These steps have been researched for the purpose of discovering the scientific uses of Vectorette PCR and how they can be applied.

Steps

Vectorette PCR is comprised of three main steps. The first step includes utilizing a restriction enzyme in order to accomplish digestion of the sample DNA. After that is completed, a Vectorette library is brought together by ligating the Vectorette units to the appropriate DNA fragments which were previously digested. Ligation is the act of binding two things together. The third and last step has two parts to it. This is due to there being two primers, the initiating primer (IP) and the Vectorette primer (VP), that act in different stages. During the first part, the IP works on amplifying the primer extension while the VP remains hybridized with the product; thus, any background amplification is not carried out at this stage. However, this changes during the last and following part of PCR as the priming that is performed comes from both the IP and the VP.

Research

A lot of research has been conducted on Vectorette PCR and the applications it has in the field of biology. Scientists used Vectorette PCR to take the transgene flanking DNA and isolate it. They used this technique on the DNA belonging to mice that was next to transgene sections. From this the scientists were able to show that the use of Vectorettes is capable of facilitating the recovery and mapping of sequences in complex genomes. They’ve also found that Vectorette PCR can help in the analysis of sequences by subvectoretting when PCR products of a large size are the subject at hand.

Other work has looked at developing a method using Vectorette PCR in order to accomplish genomic walking. By using Vectorette PCR, scientists were able to acquire single-stranded DNA which were obtained from PCR products in order to sequence them. From this an approach was identified in which the amplification of sequences which were previously uncharacterized was possible. This research demonstrates how novel sequences can be rapidly developed when only a known sequence of DNA is used to start.

Further research has experimented with the creation of a method that progresses the isolation of microsatellite repeats. By using Vectorette PCR, researchers have found a rapid technique to accomplish this with novel, microsatellite repeats. They have attempted and succeeded in using this technique to isolate an amount of six microsatellite repeats..

Vectorette PCR has also been used to not only identify genomic positions of insertion sequences (IS) but also to map them. Research on this has shed light on a way to complete the typing of microbial stains and the identification and mapping of things like IS insertion sites that reside in microbial genomes. Vectorette PCR proves useful when it comes to rapidly and simply surveying genomes’ IS elements.

There is a lot more research that has been conducted and the above information is a few of the most prominent examples. In conclusion, research on Vectorette PCR has proven to yield a plethora of results. These results have provided many uses that have come about due to Vectorette PCR.

Uses

The uses that have been derived from Vectorette PCR are many and have been useful to the science of biology. For example, it gives rise to methods that can help during the outbreaks of diseases by making it easier to subtype pathogens that are similar or closely related. Earlier in this page it was noted that Vectorette PCR can give rise to multiple functions that can be performed on novel DNA sequences located near a sequence that is already known. These functions like isolating DNA, amplifying it, and analyzing it are behind the uses for Vectorette PCR. These uses are things like genome walking, DNA sequencing for the termini of Yeast Artificial Chromosomes (YAC) and cosmid inserts, being able to map introns and promoters in genomic DNA and regions with mutations, facilitating the sequencing of clones of a large size, and filling in the gaps that arise during the mapping of genomes. Vectorette PCR also gives the user an advantage than if he/she were using other existing technologies. The user will be able to carry out tasks like gene manipulation that is cell-free, Vectorette PCR with minimal material to start with, and performing Vectorette PCR with DNA that needs not be of high purity. These advantages allow the user to save time and resources while increasing the range of DNA that can be targeted.