Gene identification and separation has helped quantify the differences in performance and several other biological aspects. It has helped scientists know why organisms of the same species do not respond in the same way when subject treatment. This is only possible when you do a PCR analysis which helps you identify differences in DNA. We have several real time PCR kit.
Real time PCR unlike conventional DNA analysis allows for detection and amplification of the strands at the exponential stage. Traditional analysis amplifies the nucleic acid at the end point. This is slow making real time have an upper hand. We have the five prime nuclease assay.
The nuclease assay has several constituents or pieces and tools brought together that make it work. First, we have the digesting polymerase enzymes. It is an enzyme and as any other polymerase enzyme it degrades the DNA strand but only at the five prime end. Such an enzyme we say has an exo-nuclease activity only and not the endonuclease activity. It works with the help of FRET which helps you transfer the energy.
FRET works with respect to energy transfer. And usually energy is transferred from a body of high energy to a body of low energy when the two bodies are in close proximity. FRET works in the presence of a probe.
Probes are tools used in the nuclease assay kit. Annealing the strands occurs at an exact specified temperature. It has the informer dye molecule with high energy sitting at the five prime end and the quencher dye molecule with low energy at the three prime energy.
At the initial stage the sequencer dye at the five prime end is in close proximity with the receiver dye. This then means that every energy emitted by the molecule is captured by the quencher at the three prime end. When the strand is excited after the probe attached to the nuclease in the 5 prime end of the strand, the distance between these two dyes in the machine increases. The upstream dye emits the energy that is captured by the sequence signal tool.
The signal detector is connected to a computer which has a software that makes it plot graphs of these amplifications. The amplifications in real time are considered when they surpass the threshold. The threshold line is drawn in the exponential phase when the reaction hits the fluorescent intensity. This is where we have very accurate reading of the cycles. The is the cycle at which the sample reaches this threshold.
Instead of using the reporter and dyes we can also use the green coloration in the SYBR kit. SYBR sequencing of the genes is the process the florescent intensity is increased as more energy is expended by the dye. It is different from the five prime nuclease assay in that it amplifies or unwinds the double strands from both ends that is the upstream and downstream end.
There is also one step kit as one of the equipments used in the sampling real time. It is applied in ribose nucleic acid (RNA)analysis. It sequences the signals of messenger RNA (mRNA) which also called the true DNA.
Real time PCR kit have helped quicken the results of the analysis in the laboratory. This results are also very precise and even the tiny differences that the conventional types would have overlooked or stack together as a thick band. This then mean that you have to be thorough in the process of extraction be it the DNA or RNA.
Real time PCR unlike conventional DNA analysis allows for detection and amplification of the strands at the exponential stage. Traditional analysis amplifies the nucleic acid at the end point. This is slow making real time have an upper hand. We have the five prime nuclease assay.
The nuclease assay has several constituents or pieces and tools brought together that make it work. First, we have the digesting polymerase enzymes. It is an enzyme and as any other polymerase enzyme it degrades the DNA strand but only at the five prime end. Such an enzyme we say has an exo-nuclease activity only and not the endonuclease activity. It works with the help of FRET which helps you transfer the energy.
FRET works with respect to energy transfer. And usually energy is transferred from a body of high energy to a body of low energy when the two bodies are in close proximity. FRET works in the presence of a probe.
Probes are tools used in the nuclease assay kit. Annealing the strands occurs at an exact specified temperature. It has the informer dye molecule with high energy sitting at the five prime end and the quencher dye molecule with low energy at the three prime energy.
At the initial stage the sequencer dye at the five prime end is in close proximity with the receiver dye. This then means that every energy emitted by the molecule is captured by the quencher at the three prime end. When the strand is excited after the probe attached to the nuclease in the 5 prime end of the strand, the distance between these two dyes in the machine increases. The upstream dye emits the energy that is captured by the sequence signal tool.
The signal detector is connected to a computer which has a software that makes it plot graphs of these amplifications. The amplifications in real time are considered when they surpass the threshold. The threshold line is drawn in the exponential phase when the reaction hits the fluorescent intensity. This is where we have very accurate reading of the cycles. The is the cycle at which the sample reaches this threshold.
Instead of using the reporter and dyes we can also use the green coloration in the SYBR kit. SYBR sequencing of the genes is the process the florescent intensity is increased as more energy is expended by the dye. It is different from the five prime nuclease assay in that it amplifies or unwinds the double strands from both ends that is the upstream and downstream end.
There is also one step kit as one of the equipments used in the sampling real time. It is applied in ribose nucleic acid (RNA)analysis. It sequences the signals of messenger RNA (mRNA) which also called the true DNA.
Real time PCR kit have helped quicken the results of the analysis in the laboratory. This results are also very precise and even the tiny differences that the conventional types would have overlooked or stack together as a thick band. This then mean that you have to be thorough in the process of extraction be it the DNA or RNA.
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