Why do we radiolabel the primer in Sanger's Method?
Why do we radiolabel the primer in Sanger's Method?
Homework Answers
The application of primer in Sanger's method is used to determine the start site of transcription. This technique involves a radiolabelled primer, which is complementary to an area near the 3' end of mRNA. The primer is permitted to anneal to the RNA and reverse transcriptase is used to synthesize cDNA from the RNA until it reaches the 5' end of the RNA. By denaturing the hybrid and using the extended primer cDNA acts as a marker on an electrophoretic gel and verify the transcriptional initiating site. It is done by the compare and based on its locality on the gel with the DNA sequence by using the same primer on the DNA template strand. The exact nucleotide by which the transcription initiates and can be identified by matching the labelled extended primer with the marker nucleotide and both share the same migration distance on the gel.
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1) Why do we apply an indirect method in iodometry? 2) What is the function of starch in this experiment and why do not we add it at the beginning of the titration? 3) What is the name of primary standard used in the standardization of thiosulfate solution? 4) What is the reason of using iodine rarely as primary standard for the standardization of thiosulfate solution?
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