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1. What are the degenerate primers in the PCR amplification protocol? What do they do? 2. Suppose you begin a PCR reaction wi
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Answer #1

Question 1

Answer: Degenerate primers are a mixture of oligonucleotides that are similar (Codes of same amino acid sequence) but not identical (different in nucleotide base composition). These primers are designed based on conversed aminoacid sequence of the protein. As each aminoacid is coded by more than one codon, these primers are designed in such way that they have all possible combination of bases that codes for same aminoacids.

Degenerate primers amplify the same gene from different organisms.

Question 2

Answer: During PCR, the amount of DNA is double after every cycle and hence the number copies of DNA after n cycle is caluculated using the formula of 2n.

If you start with a one piece of DNA. After 30 cycle of PCR, the number of pieces of double stranded DNA after 30 cycles is 230 = 1.02 x 109 copies.

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