Question

1. You perform a restriction endonuclease assay on an uncharacterized DNA molecule, using Pstl, Sall, and Xhol. When you run the DNA on a 1% agarose gel, you obtain the following information regarding fragment number and length (all lengths are in bp): Pstl (P) 700 200 Sall(s) 600 300 Xhol (X) 500 350 50 Pstl + Sall (P+S) 600 200 100 Pstl + Xhol (P+X) 500 200 150 50 Sall + Xhol (S+X) 500 250 100 50 Solve the following restriction map problem on a separate piece of paper. Fill in the possible restriction map of the DNA molecule below. Indicate locations of recognition sites and lengths of intervening DNA fragments. Assume the original DNA molecule was linear. Answer base pair (bp) length for each fragment with the corresponding letter. Indicate restriction enzyme cuts by the first letter of the enzyme (for example for Xhol just use X) for numbers 1-4. 2 3 4

Answer 1

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Ne que Here given single digestion and Sequence digested with double digestion CONA two restriction Enzymes) Pst! Pst & sall (P+ 3) 100 200 Sall 300 GOD 1 2009 200 Sall Pst 1 xhol(x) Pstt + xhol (P+x) 50. 35. 5. 500 200 1 xhol Pst 1 xhol Here be 200 would dee P & 150 here that ASO Pstl alone it the are new Care 3 fragments of 500, 350 in ↑ 150 9.50 restriction digestion site is present in between it digesting would have generated too & 200 bp DNA Sequences. Here to identify restriction digestion sile just see which fragments produced in double digestion. In thos & so be produced double digestion of Pst 1 four fragment produced of 500, 200 150 & 50 bp 500 bp & so bp fragments have been splitted into 350 bp fragment must 200 & 150 bp respiction digestion site is present & 150 bp fragment (st) are ane ahol & common ments are but newly generated it 200 150 bp frag 4 means 200 in between 500 sobp ↑ 1007 25 op so X s loobp loogp Isobo (o lo xhol oll Pst1 xhol (2) (3) (4) diagram. aboue final Answer is