Amplifying DNA: PCR cell
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1.The importance of DNA cloning:
		Current DNA technology is based on two different approaches:
		a. Specific amplification (DNA cloning) which involves cell-based DNA cloning (involving a vector/replicon and a suitable host cell) and in vitro DNA cloning (PCR )
		b. Molecular hybridization where the DNA fragment of interest is specifically detected using a mixture of different sequences
		
		
		
		2. Polymerase Chain Reaction (PCR ): features & Applications:
		Template DNA: DNA (linear or circular) or cDNA (complementary DNA produced from produced mRNA by reverse transcriptase)
		Primers: pairs of oligonucleotides each 18-25 nucleotides long; 40%-60% GC content; melting temp of both should not differ by >5oC; 3’ terminal sequences of any primer should not be to any sequences of the other primer in the pair; self-complimentary sequences (inverted repeats) of
		>3 bp should be avoided.
Cycling Nature & exponential amplification: denaturation; primer annealing; and DNA synthesis (extension).
		Regular Taq DNA polymerase lacks 3’ -> 5’ exonuclease activity needed to provide proof-reading function.
		
		
		











