300 Laboratory 18C b. Decontaminate the gel and any staining solution that will not be reused before leaving the laboratory c. Wash your hands RESULTS AND DISCUSSION Observe the photograph of your stained gel. The PCR control (unamp should have no evidence of amplified DNA product and PCR lanes should each exhibit a single fragment purified 1,100 bp. Depending upon responding to about ho far your samples have each product lane may also di a migrat we ow-molecular weight product, called \"primer dimer,\" that when ahead of the bromophenol blue marker Primer-dimer results stranded primers partially hybridize to form a short double-stranded sequence that primes extension polymerase. The primer dim may er be reduced in intensity in the purified PCR product lane Purified POR POR PCR NB+H control product product 1,100 BP Ideal Gel 1. is the purpose of the phenol/chloroform extraction? What is the purpose of the ethanol precipitation? 2. What is the purpose of the spin-column chromatography 3. Draw a schematic diagram to illustrate cycles the first three of your PCR reaction. It may be easier to use different colored pens or p 4. What would be the result of replacing the Tag polymerase E. coli DNA polymerase?
4. Unlike Taq DNA polymerase, E.coli DNA polymerase is not heat-stable and will denature during the strand denaturation step of the PCR reaction.
DNA polymerase is the chief enzyme of DNA replication. It helps to synthesize and catalyze the bonds between the nucleic acids in DNA (adenine, cytosine,uanine, thymine).DNA polymerase is an enzyme that are involved in DNA synthesis.
7. Advantages:
PCR cloning differs from traditional cloning in that the DNA fragment of interest, and even the vector can be amplified by the Polymerase Chain Reaction (PCR) and ligated together, without the use of restriction enzymes. PCR cloning is a rapid method for cloning genes, and is often used for projects that require higher throughput than traditional cloning methods can accommodate. It allows for the cloning of DNA fragments that are not available in large amounts.
Disadvantages:
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