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What are the differences between conventional pcr andreal time pcr?
PCR allows amplification of DNA for a specific gene, after too many cycles of PCR the result will reach saturation, basically meaning all of the DNA has been amplified. Conventional PCR will basically tell you whether or not a gene is expressed in your sample. This can be done semi-quantitavely if the PCR is performed for a low number of cycles, ie it will tell you whether one sample expresses more of your gene of interest than another sample. The results are seen by separating the PCR products by agarose gel/ethidium bromide electrophoresis. Real-time PCR will record exactly what cycle of PCR a detectable level of amplified product became detectable, giving a far more accurately quantifiable estimation of gene expression.
What is the difference between Clean-room and sterile-room?
You may want to double-check this, but I believe PCR-clean simply means that there are no DNases or RNases on the item, but they still could have nucleic acid on them. Essentially there is nothing on them that would interfere with nucleic acid amplification achieved with PCR, but any genetic material they may have will be amplified. Sterile means that there is absolutely no genetic material on the item itself (usually achieved via autoclaving where the temperatures climb so high that they would denature the DNases and RNases anyway). Nutshell: PCR-clean = wiping with RNA away and DNA away Sterile = bleaching and autoclaving
Why 10x buffer use in pcr?
It provides a suitable chemical environment for optimum activity and stability of the DNA polymerase.
What are the steps in a direct-sequencing experiment?
Lyse cells, purify DNA, amplify genes by PCR, and insert genes into plasmid
Why are scientists so interested in extremophiles?
Extreme environments have been useful to scientists in inventing PCR. It was in an extreme environment like the geysers of Yellowstone that a scientist discovered that a bacteria was living in the extremely hot water and yet still could function. Before PCR we knew we could separate a strand of DNA by heating it, but there was no polymerase to duplicate it that would work at such a high temperature. The bacteria in the hot water had a polymerase that would. So now scientists use that to do PCR and create many copies of DNA.
What is the use of dNTP?
The use of dNTP is PCR and multiplex PCR
Whats the purpose of a multiplex PCR?
Multiplex PCR allows for the amplification of multiple target DNA sequences in a single reaction. This method increases efficiency and throughput by reducing the number of reactions needed. It is commonly used for genotyping, pathogen detection, and gene expression analysis.
What is pcr and types of pcr?
PCR (polymerase chain reaction) is a molecular biology technique used to amplify a specific segment of DNA. There are various types of PCR, including quantitative PCR (qPCR) for quantification of DNA, reverse transcription PCR (RT-PCR) to amplify RNA, nested PCR for increased specificity, and digital PCR for absolute quantification of nucleic acids.
What is the difference between a PCR tire and a TBR tire?
PCR: Passenger Car Radial TBR: Truck and Bus Radial A TBR tire can handle a heavier load than a PCR tire, and it's usually bigger.
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)
Differentiate between quantitative and real time PCR?
: Differentiate between quantitative and real time PCR.
What is the difference between touch down and gradient PCR?
Touch-down PCR is a method where the annealing temperature decreases in each cycle to increase specificity, while gradient PCR involves testing a range of annealing temperatures in a single experiment to determine the optimal temperature for PCR amplification. Touch-down PCR is useful for reducing nonspecific amplification, while gradient PCR is helpful for identifying the optimal annealing temperature for a specific primer pair.
What is the defference between Real-time PCR and reverse transcriptase PCR?
Real-time PCR is a technique used for quantifying DNA in real-time during the PCR process, while reverse transcriptase PCR (RT-PCR) is used to detect RNA by first converting it to complementary DNA (cDNA) using reverse transcriptase enzyme before proceeding with PCR amplification. Real-time PCR allows for monitoring the amplification process as it occurs, while RT-PCR is specifically used for analyzing RNA levels.
What are the different types of polymerase chain reaction techniques?
types of pcr: AFLP -PCR. Allele-specific PCR. Alu-PCR. Assembly -PCR. Assemetric -PCR. Colony -PCR. Helicase dependent amplification. Hot start pCR. Inverse -PCR. Insitu -pCR. ISSR-PCR. RT-PCR(REVERSE TARNSCRIPTASE). REAL TIME -PCR
What virus can be detected by polymerase chain reaction?
Polymerase chain reaction (PCR) can detect a wide range of viruses, including influenza, HIV, hepatitis C, herpes simplex virus, cytomegalovirus, and SARS-CoV-2. PCR is a sensitive and specific method for amplifying and detecting viral genetic material in clinical samples.
What is the enzyme that synthesized DNA used in PCR that distinguishes it from the equivalent enzyme that carry out the same function in our cells or those of most bacteria?
The enzyme used in PCR to synthesize DNA is called DNA polymerase. The key difference is that the DNA polymerase used in PCR, such as Taq polymerase, is derived from a thermophilic bacterium called Thermus aquaticus and can withstand the high temperatures used in the PCR cycling process. This distinguishes it from the equivalent enzyme in our cells or most bacteria, which would be denatured by the high temperatures of PCR.
Does 50ul PCR product work better than 25ul PCR product?
The volume of PCR product used does not necessarily determine its effectiveness. The critical factors that affect PCR performance include the quality and concentration of the DNA template, presence of inhibitors, primer design, and PCR conditions. It is best to optimize these parameters for successful PCR amplification rather than focusing solely on the volume of PCR product.
