Taq polymerase

ABSTRACT Taq Polymerase is essential in Polymerase Chain Reaction(PCR) experiments to obtain a PCR amplification of an unknown gene. The unknown gene is then ligated into a vector plasmid, which is placed in a bacterium Escherichia Coli to transform and multiply. Ultimately, identification and characterization of the unknown gene is done using electrophoreses and gel imaging. Cloning techniques such as the one performed have been used for many years to isolate genes from a variety of species. Isolation

using SSR markers. The PCR reactions were performed with 15 μl final volume containing 50 ng (2 µl) of genomic DNA, 1.5 µl of 10X buffer containing 25mM MgCl2, 1.5 µl of 2.5 mM dNTP mix, 1 µl each of forward and reverse primers (5 mM), and 1U of Taq polymerase. The

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enveloped in the master mix, that consists of dNTPs, a buffer, primers, and Taq DNA polymerase. The dNTPs are deoxyribonucleoside triphosphate which are basically the nucleotides needed to elongate a DNA sequence. The buffer is important because it creates the correct pH and salt condition for DNA polymerase and because it also harbors magnesium ions in it, which serve as a cofactor for DNA polymerase. Taq DNA polymerase is vital because it is a heat stable enzyme derived from the bacteria Thermus

Taq polymerase does not work on RNA samples, so PCR cannot be used to directly amplify RNA molecules. The incorporation of the enzyme reverse transcriptase (RT), however, can be combined with traditional PCR to allow for the amplification of RNA molecules. Afterwe add our RNA sample to the PCR machine, add a DNA primer as usual and allow it to anneal to your target molecule. Then add RT along with dNTPs, which will elongate the DNA primer and make a cDNA copy of the RNA molecules and run the PRC

PCR (Polymerase Chain Reaction) is a laboratory technique that takes specific fragments of DNA and amplifies them using primers and a polymerase that can withstand high temperatures. The materials needed to complete this experiment include the DNA fragments that are to be amplified, two primers (one to attach to the top strand of the DNA fragment of interest and one to attach to the bottom strand on the other end), Taq DNA polymerase, dNTP mix, MgCl2, PCR buffer and the PCR machine. Primers are necessary

Using PCR and Gel Electrophoresis to Determine Genotype In certain situations, it is necessary to identify DNA retreived from a sample. When there is a small sample in need of identification, Polymerase Chain Reactions are used to multiply the DNA in the sample in to many identical samples. The DNA retrieved from the reaction can then be imported into an aparatus using gel electrophoresis to compare the sample of DNA to other samples. In our experiment we learned the how to replicate tiny

The purpose of this experiment was to amplify the α-amylase gene in Bacillus subtilus using a polymerase chain reaction. Bacillus subtilus is a gram-positive bacterium that resides in soil and the gastrointestinal tract of animals and primarily uses aerobic respiration for survival (Yu et al., 2015). Due to it’s ability to take part in the fermentation process, B. subtilus can produce large amounts of the α-amylase enzyme which is responsible for catalyzing the digestion of polysaccharides, such

Polymerase chain reaction, also known as PCR, is a technique to make multiple copies of a specific DNA region. PCR relies on a thermostable DNA polymerase and requires DNA primers for the specific regions of DNA that is being used. The main goal of PCR is to make enough of the specific DNA, in that region, that can be analyzed for many different reasons. For example, having enough of the DNA amount for forensic scientist would help them match crime scene DNA to the actual suspect. PCR requires

Polymerase Chain Reaction (PCR) is a process that uses primers to amplify specific cloned or genomic DNA sequences with the help of a very unique enzyme. In this experiment, PCR was used to amplify the cDNA inserts from three purified DNA plasmids, taking into consideration that the cDNAs were all different in size, as predicted in the previous study, so there would be different sized DNA bands on agarose gel as well. In PCR, a primer and dNTPs are added along with a DNA template and the Taq DNA