TUGAS REKGEN Restriction Enzymes

Restriction Enzymes

Directions: Identify the restriction sites for each of the examples given. Show the cuts , sticky ends or blunt, number of DNA fragments produced and the number of base pairs in each (count the top row). (If there are three nucleotides on either side of the dash it is a blunt cut. If there are fewer than three bases on either side of the dash it produces “sticky ends” cut.)

Remember:  Analyze in the 5’à 3’ direction

1.  HindIII--- 5' GTC ↓GAC 3'

5' ACGACGTAGTC↓GACTTATTAT GTC↓GACCCGCCGCGTGTC↓GACCATCA 3'

3' TGCTGCATCAG↓CTGAATAATACAG↓CTGGGCGGCGCACAG↓CTGGTAGT 5'

Tipe pemotongan ujung tumpul (Blunt ends), jumlah hasil pemotongan fragmen DNA sebanyak 4.

Jumlah pasangan basa nitrogennya adalah 11 bp, 13 bp, 15 bp, dan 8 bp.

2.  EcoRI --- 5' G ↓AATTC 3'

5' ACG ACGTATTAG↓AATTCTTAT CCGCCGCCGG↓AATTCT CATCA 3'

3' TGC TGCATAATCTTAA↓GAATAGGCGGCGGCCTTAA↓GAGTAGT 5'

Tipe pemotongan ujung lancip (Sticky ends), jumlah hasil pemotongan fragmen DNA yaitu sebanyak 3.

Jumlah pasangan basa nitrogen nya adalah 12 bp, 19 bp, dan 11 bp.

3.  HaeIII --- 5' CC ↓ GG 3'

5' ACGCC ↓GGCCGTATTAT CC ↓GGATCCGCCG CC ↓GGCTGTCCC ↓GGATCA 3'

3' TGCGG ↓CCGGCATAATAGG ↓CCTAGGCGGCGG ↓CCGACAGGG ↓CCTAGT 5'

Tipe pemotongan ujung tumpul (Blunt ends), jumlah hasil pemotongan fragmen DNA sebanyak 5.

Jumlah pasangan basa nitrogen nya adalah 5 bp, 13 bp, 12 bp, 9 bp, dan 6 bp.

4.  BamI --- 5' CCTAG ↓G 3'

5' ACGCCTAG ↓GACGTATTATCCTAG ↓GTAT CCGCCGCCGT CATCA 3'

3' TGCG ↓GATCCTGCATAATAG ↓GATCCATAGGCGGCGGCAGTAGT 5'

Tipe pemotongan ujung lancip (Sticky ends), jumlah hasil pemotongan fragmen DNA yaitu sebanyak 3.

Jumlah pasangan basa nitrogen nya adalah 8 bp, 15 bp, dan 19 bp.

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

5' ACGGTC↓GACACGTATTATTAGTC↓GACTCCGCCGCCGCC↓GGTCATCA 3'

3' TGCCAG↓CTGTGCATAATAATCAG↓CTGAGGCGGCGGCGG↓CCAGTAGT 5'

Tipe pemotongan ujung tumpul (Blunt ends), jumlah hasil pemotongan fragmen DNA yaitu sebanyak 4.

Jumlah pasangan basa nitrogen nya adalah 6 bp, 17 bp, 15 bp, dan 8 bp.

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

5' ACGCC↓GGACGTACCTAGGTTTAGTC↓GACTC CGCCG CCCCTAG↓GGTCATCA 3'

3' TGCGG↓CCTGCATGGATCCAAATCAG↓CTGAGGCGGCGGG↓GATCCCAGTAGT 5'

Tipe pemotongan ujung lancip (Sticky ends), jumlah hasil pemotongan fragmen DNA yaitu sebanyak 4.

Jumlah pasangan basa nitrogen nya adalah 5 bp, 20 bp, 17 bp, dan 8 bp.

by: Nanda Dwi Martina & Siti Apriliyanti Khadijah (Biologi B)