Question

Using PCR an RFLP was identified on the first chromosome of wild strains of Banana slugs isolated in Santa Cruz. In strain C, the fragment could be cut with EcoR1 yielding 3 kb and 4 kb bands when run on a gel. In strain E, the fragment was not cut by Eco1 resulting in a 7 kb band run on a gel. A strain E in lividual that is also homozygous for the first chron come recessive mutation day-glo (dg) (bright yellow) is discovered and crossed to strain C individual that is homozygous for the dominant normal allele of Day-glo (Dg) (green). The resulting F1 individuals from this cross are mated with strain E individuals homozygous for the dg recessive mutation (and the RFLP) and the F2 progeny are scored for the body color and PCR bands on the gel for EcoR1 digestion. Four classes of progeny are produced: class 1:51 bright yellow individuals that produce a 7 kb band on the gel; class 2: 15 bright yellow individuals that produce 7kb, 3 kb and 4 kb bands on the gel: class 3: 52 green individuals that produce 7kb, 3kb and 4kb bands; class 4: 18 green individuals that produce a 7 kb band on the gel.

Answer 1

PCR is polymerase chain reaction which is used for the amplification of a DNA molecule. In this technique, following are the most important reagents which are required -

Pair of primers - Primers bind to 3' end of single stranded DNA molecule and provide free hydroxyl group for taq polymerase to add deoxyribonucleotides

Taq polymerase - this is the enzyme which helps in the polymerisation of daughter DNA molecule

MgCl2 - this provides magnesium Ion which are the cofactor for taq polymerase enzyme

Template DNA - which act as the parental DNA strand against which complementary daughter DNA molecule is synthesized

dNTPs - single molecules of deoxyribonucleotides are added by tag polymerase enzyme to the growing daughter DNA molecules

Buffer - buffer is used in this process to maintain a constant during the process of amplification

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ECOR1 / ELOR I SY7 (c) 14 (8) incut strain c .. strain & 7 TM

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