Question

1. Directions: Identify the restriction sites for each of the examples given. Show the cuts, mention if they are sticky ends or blunt, 5’ overhangs or 3 ‘overhangs and the number of DNA fragments produced and the number of base pairs in each (count the top row).

            Remember: Analyze in the 5’à 3’ direction

1. HindII --- 5' GTC ↓GAC 3'

5' ACGACGTAGTCGACTTATTAT GTCGACCCGCCGCGTGTCGACCATCA 3'

3' TGCTGCATCAGCTGAATAATACAGCTGGGCGGCGCACAGCTGGTAGT 5'

Number of pieces of DNA _______

2. EcoRI --- 5' G ↓AATTC 3'

5' ACG ACGTATTAGAATTCTTAT CCGCCGCCGGAATTCT CATCA 3'

3' TGC TGCATAATCTTAAGAATAGGCGGCGGCCTTAAGAGTAGT 5'

Number of pieces of DNA _______

3. HaeIII --- 5' CC ↓ GG 3'

5' ACGCCGGCCGTATTAT CCGGATCCGCCG CCGGCTGTCCCGGATCA 3'

3' TGCGGCCGGCATAATAGGCCTAGGCGGCGGCCGACAGGGCCTAGT 5'

Number of pieces of DNA _______

4. BamI --- 5' CCTAG ↓G 3'

5' ACGCCTAGGACGTATTATCCTAGGTAT CCGCCGCCGT CATCA 3'

3' TGCGGATCCTGCATAATAGGATCCATAGGCGGCGGCAGTAGT 5'

Number of pieces of DNA _______

5. HindII --- 5' GTC ↓- GAC 3' and HaeIII --- 5' CC ↓ GG 3'

5' ACGGTCGACACGTATTATTAGTCGACTCCGCCGCCGCCGGTCATCA 3'

3' TGCCAGCTGTGCATAATAATCAGCTGAGGCGGCGGCGGCCAGTAGT 5'

Number of pieces of DNA _______

6.   HindII --- 5' GTC ↓ GAC 3', HaeIII --- 5' CC ↓ GG 3' and BamI --- 5' CCTAG ↓ G 3'

5' ACGCCGGACGTACCTAGGTTTAGTCGACTC CGCCG CCCCTAGGGTCATCA 3'

3' TGCGGCCTGCATGGATCCAAATCAGCTGAGGCGGCGGGGATCCCAGTAGT 5'

Number of pieces of DNA _______

Answer 1

1. HindII --- 5' GTC ↓GAC 3'
5' ACGAC"GTA-↓-GTC"GACTTATTAT"GTC-↓-GAC"CCGCCGCGT"GTC-↓-GAC"CATCA 3'
3' TGCTG"CAT-↓-CAG"CTGAATAATA"CAG-↓-CTG"GGCGGCGCA"CAG-↓-CTG"GTAGT 5'
Number of pieces of DNA : Four(4)
Type of the end: Blunt

2. EcoRI --- 5' G ↓AATTC 3'
5' ACGACGTATTA"G-↓-AATTC"TTATCCGCCGCCG"G-↓-AATTC"TCATCA 3'
3' TGCTGCATAAT"CTTAA-↓-G"AATAGGCGGCGGC"CTTAA-↓-G"AGTAGT 5'
Number of pieces of DNA : Three(3)
Type of the end: hanging: 5' hanging ends

3. HaeIII --- 5' CC ↓ GG 3'
5' ACG"CC-↓-GG"CCGTATTAT"CC-↓-GG"ATCCGCCG"CC-↓-GG"CTGTC"CC-↓-GG"ATCA 3'
3' TGC"GG-↓-CC"GGCATAATA"GG-↓-CC"TAGGCGGC"GG-↓-CC"GACAG"GG-↓-CC"TAGT 5'
Number of pieces of DNA: Five(5)
Type of the end: Blunt ends

4. BamI --- 5' CCTAG ↓G 3'
5' ACG"CCTAG-↓-G"ACGTATTAT"CCTAG-↓-G"TATCCGCCGCCGTCATCA 3'
3' TGC"G-↓-GATCC"TGCATAATA"G-↓-GATCC"ATAGGCGGCGGCAGTAGT 5'
Number of pieces of DNA : Three (3)
Type of the end: hanging: 3' hanging ends

5.  
5' ACG"GTC-↓-GAC"ACGTATTATTA"GTC-↓-GAC"TCCGCCGCCG"CC-↓-GG"TCATCA 3'
3' TGC"CAG-↓-CTG"TGCATAATAAT"CAG-↓-CTG"AGGCGGCGGC"GG-↓-CC"AGTAGT 5'
Number of pieces of DNA : Four (4)
Type of the end: Blunt ends

6. HindII --- 5' GTC ↓ GAC 3', HaeIII --- 5' CC ↓ GG 3' and BamI --- 5' CCTAG ↓ G 3'
5' ACG"CC-↓-GG"ACGTA"CCTAG-↓-G"TTTA"GTC-↓-GAC"TCCGCCGCC"CCTAG-↓-G"GTCATCA 3'
3' TGC"GG-↓-CC"TGCAT"G-↓-GATCC"AAAT"CAG-↓-CTG"AGGCGGCGG"G-↓-GATCC"CAGTAGT 5'
Number of pieces of DNA : Five(5)
Type of the end: hangin: 3' hanging and Blunt ends