1.State reasons why primers are phosphorylated at their 5’ end during synthesis
ID: 262928 • Letter: 1
Question
1.State reasons why primers are phosphorylated at their 5’ end during synthesis reaction
2.What are primers ? explain in details with examples
3.During site directed mutagenesis it is important that the host cell used is deficient in mis match repair (mutS mutants) in the first transformation but not necessary at the second transformation. Give reasons for this.
4.In site directed mutagenesis why is digestion of restriction enzymes such as NdeI necessary prior to the second transformation ?
5.How are primers designed for site directed mutagenesis ? and what features of the primers are essential when they are been compared?
Explanation / Answer
1. Primers are phosphorylated at their 5’ end during synthesis reaction because they are used for site-directed mutagenesis.
2. Primer is a short segment of DNA that range from 18 base pair to 22 base pair in length. They are initiator of DNA synthesis. DNA polymerase is the enzyme responsible for the replication, does not initiate the DNA synthesis. It is initiated with the help of primer. Primer is generally used in polymerase chain reaction to produce multiple copies of the gene. The two types of primer are forward and reverse primer. In order to design primer, it length should be between 18–30 nucleotides. Melting point temperature should be between 65°C and 75°C. GC content should be between 40-60%.
EXmaple of primer is 5' atgcgtccggcgtagag 3' .
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