We have four DNA sequences that we need to digest (cut up), and have 6 restricti

Question

We have four DNA sequences that we need to digest (cut up), and have 6 restriction enzymes in stock in our lab. We want to find out which restriction enzyme we can use on each sample.

For each sequence, please indicate which restriction enzyme(s) would cut within each sequence. Each sequence may be cut by one, more than one, or no restriction enzymes.

Following the sequences are two questions. Please choose from the same list of enzymes to indicate the correct answer to these questions.

5' G|AATTC 3'

3' CTTAA|G 5'

5' A|AGCTT 3'

3' TTCGA|A 5'

5' G|GATCC 3'

3' CCTAG|G 5

5' AG|CT 3'

3' TC|GA 5'

5' CCC|GGG 3'

3' GGG|CCC 5'

5' GCG|C 3'

3' C|GCG 5'

(The | symbol indicates the specific cut sites.)

Each answer may be used once, more than once, or not at all.

5' -TAGACTGAATTCAAGTCA- 3'

3' -ATCTGACTTAAGTTCAGT- 5'

5' -ATACGCCCGGGTTCTAAA- 3'

3' -TATGCGGGCCCAAGATTT- 5'

5' -CAGGATCGAAGCTTATGC- 3'

3' -GTCCTAGCTTCGAATACG- 5'

5' -AATAGAATTCCGATCCGA- 3'

3' -TTATCTTAAGGCTAGGCT- 5'

Which restriction enzyme(s) is(are) 4-cutters (i.e. they recognize and cut at 4-bp sequences, instead of 6-bp sequences)?

Which restriction enzyme(s) do not leave a "sticky end" after cutting the DNA?

Alu

AluI, SmaI

All 6 of these restriction enzymes.

None of the 6 restriction enzymes listed.

BamH1

HindIII, AluI

EcoR1

SmaI

SmaI and HbaI

HbaI

HindIII

AluI, HbaI

EcoRI, AluI

AluI

BamHI, AluI

Restriction enzyme Digestion pattern EcoRI

5' G|AATTC 3'

3' CTTAA|G 5'

HindIII

5' A|AGCTT 3'

3' TTCGA|A 5'

BamHI

5' G|GATCC 3'

3' CCTAG|G 5

AluI

5' AG|CT 3'

3' TC|GA 5'

SmaI

5' CCC|GGG 3'

3' GGG|CCC 5'

HbaI

5' GCG|C 3'

3' C|GCG 5'

Explanation / Answer

1, Ans G- ECOR1

5' -TAGACTGAATTCAAGTCA- 3'

3' -ATCTGACTTAAGTTCAGT- 5'

2. Ans H SmaI

5' -ATACGCCCGGGTTCTAAA- 3'

3' -TATGCGGGCCCAAGATTT- 5'

3. ans F HindIII, AluI

HindIII

5' -CAGGATCGAAGCTTATGC- 3'

3' -GTCCTAGCTTCGAATACG- 5'

AluI

5' -CAGGATCGAAGCTTATGC- 3'

3' -GTCCTAGCTTCGAATACG- 5'

4. Ans G.ECORI

5' -AATAGAATTCCGATCCGA- 3'

3' -TTATCTTAAGGCTAGGCT- 5'

5. 4 cutter enzymes are AluI and HbaI, Ans L

6. AluI and SmaI, Ans B

AluI-

5' AG 3' //  5' CT 3'  

3' TC 5' //  3' GA 5'

SmaI-  

5' CCC 3' // 5' GGG 3'

3' GGG 5' // 3' CCC 5'

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