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In order to calculate the concentration of DNA in PCR products (usually expressed in micrograms permicroliter), one had to first establish a standard curve that correlates the concentration of DNA and its absorbency at 280 nm. This standard graph can be set up by preparing serial dilutions of DNA of known concentration and then measuring the absorbency of the sample at 280nm. Ideally, a linear graph is seen. Now that a standard graph has been established, the product obtained at the end of a PCR reaction can be sampled for absorbency measurement. Using the absorbency value, one can estimate the concentration of DNA be interpolating on the standard graph. There are however, several calculations that that to be made in order to arrive at the final answer.
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What is the purpose of the buffer in PCR?
The purpose of the buffer in PCR, I assume you talking about the 5 or 10 times PCR buffer that is provided with enzyme. Buffer is needed to give the correct pH and pottasium ion concentration for the DNA polymerase enzyme (usually DNA polymerase from Thermus aquaticus) to function.
What controls which DNA is amplified in PCR?
The choice of primers controls which DNA is amplified in PCR.
What is difference between Qualitative PCR and Quatitative PCR?
In qualitative PCR specific DNA fragment is detected while in quantitative PCR our target DNA sequence not only is detected but its amount is determined (after reaction we can calculate the amount of DNA we had in our sample)
What are the rt pcr?
Real time PCR is uded to amplify and simultaneously quantify a targeted DNA molecule.
In a pcr reaction of DNA are first separated by?
Heating
Why pcr products are precipitated before sequencing?
The PCR product are precipitated before sequencing to increase the concentration of tamplet DNA.
How does a DNA molecule act as a template?
Template DNA is a DNA you want to amplify. So you should know what you are amplifying before a PCR or you can make it by sequencing your PCR product.
What is the sequence of the template DNA?
What do you really want to ask? template DNA is a DNA you want to amplify. So you should know what you are amplifying before a PCR or you can make it by sequencing your PCR product.
What is the purpose of the buffer in PCR?
The purpose of the buffer in PCR, I assume you talking about the 5 or 10 times PCR buffer that is provided with enzyme. Buffer is needed to give the correct pH and pottasium ion concentration for the DNA polymerase enzyme (usually DNA polymerase from Thermus aquaticus) to function.
What controls which DNA is amplified in PCR?
The choice of primers controls which DNA is amplified in PCR.
Which is used to copy DNA for DNA fingerprinting?
PCR
What is difference between Qualitative PCR and Quatitative PCR?
In qualitative PCR specific DNA fragment is detected while in quantitative PCR our target DNA sequence not only is detected but its amount is determined (after reaction we can calculate the amount of DNA we had in our sample)
What is TA cloning vector?
TA Cloning is one of the most popular methods of cloning the amplified PCR product using Taq and other polymerases. These polymerases lack 5'-3' proofreading activity and are capable of adding adenosine triphosphate residue to the 3' ends of the double stranded PCR product. Such PCR amplified product can be cloned in a linearized vector with complementary 3' T overhangs. TA cloning is brought about by the terminal transferase activity of certain type of DNA polymerase such as the Taq polymerase. This enzyme adds a single, 3'-A overhang to each end of the PCR product. As a result, the PCR product can be directly cloned into a linearized cloning vector that have single base 3'-T overhangs on each end. Such vectors are called T- vectors. The PCR product with A overhang, is mixed with this vector in high proportion. The complementary overhangs of a "T" vector and the PCR product hybridize. The result is a recombinant DNA, the recombination being brought about by DNA ligase.
What is pcr and types of pcr?
PCR is a biotechnological method to amplify your gene (DNA) of your interest. It produce millions of your DNA fragments hence used in cloning. There are variants of this method using the same thermocycling principle such as touch down PCR, gradient PCR, RFLP, multiplex PCR, Q PCR, RT PCR and so on.
Conclusion of pcr amplification?
Since the point of PCR is to amplify a copy of DNA, it would result in many copies of DNA that you want to study.
Why is MgCl2 a variable in PCR?
Mg2+ complexes with the single stranded DNA that is to be amplified, and becomes the substrate of DNA polymerase. In other words, it helps in the binding of primer (and the subsequent target DNA) to the template DNA. Different volume of Mg2+ exert different complex-forming capabilities, and thus affects the end product of PCR.
How is DNA tested?
electrophoresis,PCR
