What would you expect the average size of a restriction

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Reference no: EM131721752

You should also solve and answer the following problems:

1. HindIII and BamHI restriction enzymes have a 6 base pair recognition site. How many times would you expect them to cut in a DNA sequence of random base composition of 5000 base pairs? How many times would you expect an enzyme with a 4 by recognition site to cut the same DNA?

2. Chargaff (1955) reported the base compositions of the chromosomes of several bacteria including:

Escherichia coli 22% G 22% C 23% A 28% T
Streptomyces coelicolor 39% G 39% C 11% A 11% T

The restriction enzyme EcoRI cleaves DNA at the recognition sequence GAATTC. What would you expect the average size of a restriction fragment to be when (a) E. colt, and (b) S. coelicolor DNA is digested with EcoRI ?

3. Restriction fragments obtained upon digestion of circular plasmids are shown below (all sizes are in kb). Deduce the restriction map for each plasmid.

(a). pBI01:

EcoRI	1	1.7	2.3
BamHI	0.8	4.2
HindIII	5
EcoRI + BamHI	0.3	0.8	1	1.2	1.7
EcoRI + HindIII	0.5	1	1.2	2.3
BamHI + HindIII	0.8	1.8	2.4

(b). pBIO2:

EcoRI	0.4	0.6	5
BamHI	2.5	3.5
HindIII	6
EcoRI + BamHI	0.4	0.6	1	1.5	2.5
EcoRI + HindIII	0.4	0.6	2.2	2.8
BamHI + HindIII	1.2	1.3	3.5

EcoRI	1.3	8
BamHI	1.7	3	4.6
HindIII	0.8	8.5
EcoRI + BamHI	1	1.3	1.7	2.3	3
EcoRI + HindIII	0.8	1.3	2.7	4.5
BamHI + HindIII	0.5	0.8	1.7	1.7	4.6
triple digest:	0.5	0.8	1	1.3	1.7	1.7	2.3

4. You have radioactively labelled a linear piece of double-stranded DNA at the 5' ends. Digestion of this molecule with either EcoRI or BansHI yields the following fragments:

EcoRI:	2.8*	4.6*	6.2	7.4	8	kb
BamHI:	6.0*	10	13.0*	kb

Asterisks denote fragments that are labelled.

If unlabelled DNA is digested with both enzymes simultaneously the following fragments appear:

2.2

2.8

3.2

4.6

4.8

5.2

6.2

Deduce the restriction maps for the two enzymes.

Reference no: EM131721752

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