Restriction Mapping Example Problems RFLP Analysis Restriction Fragment

Restriction Mapping Example Problems

RFLP Analysis Restriction Fragment Length Polymorphisms Restriction enzymes from bacteria cut DNA molecules to create DNA fragments of varying lengths Separate fragments in gel electrophoresis DNA fingerprinting (CSI, paternity)

Example #1: Linear DNA Sample Fragment Lengths (bp) Uncut DNA 10, 000 DNA cut with Eco. RI 8, 000 2, 000 DNA cut with Bam. HI 5, 000 DNA cut with Eco. RI + Bam. HI 5, 000 3, 000 2, 000

Example #1: Linear DNA Sample Fragment Lengths (bp) Uncut DNA 10, 000 DNA cut with Eco. RI 8, 000 2, 000 DNA cut with Bam. HI 5, 000 DNA cut with Eco. RI + Bam. HI 5, 000 3, 000 Uncut 10, 000 bp 2, 000

Example #1: Linear DNA Sample Fragment Lengths (bp) Uncut DNA 10, 000 DNA cut with Eco. RI 8, 000 2, 000 DNA cut with Bam. HI 5, 000 DNA cut with Eco. RI + Bam. HI 5, 000 3, 000 Uncut 10, 000 bp Eco. RI 8, 000 bp 2, 000 bp

Example #1: Linear DNA Sample Fragment Lengths (bp) Uncut DNA 10, 000 DNA cut with Eco. RI 8, 000 2, 000 DNA cut with Bam. HI 5, 000 DNA cut with Eco. RI + Bam. HI 5, 000 3, 000 Uncut 10, 000 bp Eco. RI 8, 000 bp Bam. HI 5, 000 bp 2, 000 bp 5, 000 bp

Example #1: Linear DNA Sample Fragment Lengths (bp) Uncut DNA 10, 000 DNA cut with Eco. RI 8, 000 2, 000 DNA cut with Bam. HI 5, 000 DNA cut with Eco. RI + Bam. HI 5, 000 3, 000 Uncut 10, 000 bp Eco. RI 8, 000 bp Bam. HI 5, 000 bp E+B 5, 000 bp 2, 000 bp 5, 000 bp 3, 000 bp 2, 000 bp B E

Example #2: Plasmid (circular) DNA Sample Fragment Length (bp) Uncut DNA 17, 000 DNA cut with Sma. I 17, 000 DNA cut with Hae. III 17, 000 DNA cut with Pst. I 10, 000 7, 000 DNA cut with Sma. I + Hae. III 12, 000 5, 000 DNA cut with Sma. I + Pst. I 10, 000 6, 000 DNA cut with Hae. III + Pst. I 7, 000 6, 000 1, 000 4, 000

Example #2: Plasmid (circular) DNA Sample Fragment Length (bp) Uncut DNA 17, 000 DNA cut with Sma. I 17, 000 DNA cut with Hae. III 17, 000 DNA cut with Pst. I 10, 000 7, 000 DNA cut with Sma. I + Hae. III 12, 000 5, 000 DNA cut with Sma. I + Pst. I 10, 000 6, 000 DNA cut with Hae. III + Pst. I 7, 000 Uncut 17, 000 6, 000 1, 000 4, 000

Example #2: Plasmid (circular) DNA Sample Fragment Length (bp) Uncut DNA 17, 000 DNA cut with Sma. I 17, 000 DNA cut with Hae. III 17, 000 DNA cut with Pst. I 10, 000 7, 000 DNA cut with Sma. I + Hae. III 12, 000 5, 000 DNA cut with Sma. I + Pst. I 10, 000 6, 000 DNA cut with Hae. III + Pst. I 7, 000 Uncut 17, 000 6, 000 Sma. I 17, 000 1, 000 4, 000 S

Example #2: Plasmid (circular) DNA Sample Fragment Length (bp) Uncut DNA 17, 000 DNA cut with Sma. I 17, 000 DNA cut with Hae. III 17, 000 DNA cut with Pst. I 10, 000 7, 000 DNA cut with Sma. I + Hae. III 12, 000 5, 000 DNA cut with Sma. I + Pst. I 10, 000 6, 000 DNA cut with Hae. III + Pst. I 7, 000 Uncut 17, 000 Sma. I 6, 000 17, 000 1, 000 4, 000 S H Hae. III 17, 000

Example #2: Plasmid (circular) DNA Sample Fragment Length (bp) Uncut DNA 17, 000 DNA cut with Sma. I 17, 000 DNA cut with Hae. III 17, 000 DNA cut with Pst. I 10, 000 7, 000 DNA cut with Sma. I + Hae. III 12, 000 5, 000 DNA cut with Sma. I + Pst. I 10, 000 6, 000 DNA cut with Hae. III + Pst. I 7, 000 Uncut 17, 000 Sma. I 6, 000 17, 000 1, 000 S Hae. III H Pst. I 4, 000 10, 000 17, 000 P

Example #2: Plasmid (circular) DNA Sample Fragment Length (bp) Uncut DNA 17, 000 DNA cut with Sma. I 17, 000 DNA cut with Hae. III 17, 000 DNA cut with Pst. I 10, 000 7, 000 DNA cut with Sma. I + Hae. III 12, 000 5, 000 DNA cut with Sma. I + Pst. I 10, 000 6, 000 DNA cut with Hae. III + Pst. I 7, 000 S+H 17, 000 Sma. I 6, 000 H 17, 000 1, 000 4, 000 S Pst. I 10, 000 H 5, 000 12, 000 Uncut S Hae. III 17, 000 P

Example #2: Plasmid (circular) DNA Sample Fragment Length (bp) Uncut DNA 17, 000 DNA cut with Sma. I 17, 000 DNA cut with Hae. III 17, 000 DNA cut with Pst. I 10, 000 7, 000 DNA cut with Sma. I + Hae. III 12, 000 5, 000 DNA cut with Sma. I + Pst. I 10, 000 6, 000 DNA cut with Hae. III + Pst. I 7, 000 H S+H 1, 0 P 00 S S+P S 6, 000 10, 000 17, 000 Sma. I 6, 000 17, 000 1, 000 4, 000 P Hae. III H S 17, 000 P Pst. I 10, 000 5, 000 12, 000 Uncut 7, 000 P

Example #2: Plasmid (circular) DNA Sample Fragment Length (bp) Uncut DNA 17, 000 DNA cut with Sma. I 17, 000 DNA cut with Hae. III 17, 000 DNA cut with Pst. I 10, 000 7, 000 DNA cut with Sma. I + Hae. III 12, 000 5, 000 DNA cut with Sma. I + Pst. I 10, 000 6, 000 DNA cut with Hae. III + Pst. I 7, 000 S S+H H 1, 0 P 6, 000 S 6, 000 10, 000 6, 000 4, 000 H S+P Sma. I 00 12, 000 17, 000 H+P 5, 000 P Uncut 17, 000 1, 000 4, 000 S 7, 000 P 17, 000 P Pst. I 10, 000 P Hae. III H 7, 000 P

Example #2: Plasmid (circular) DNA Sample Fragment Length (bp) Uncut DNA 17, 000 DNA cut with Sma. I 17, 000 DNA cut with Hae. III 17, 000 DNA cut with Pst. I 10, 000 7, 000 DNA cut with Sma. I + Hae. III 12, 000 5, 000 DNA cut with Sma. I + Pst. I 10, 000 6, 000 DNA cut with Hae. III + Pst. I 7, 000 00 1, 0 6, 000 S 6, 000 P 17, 000 S P Pst. I 4, 000 7, 000 10, 000 P Hae. III H P 7, 000 P P 4, 000 H 6, 000 00 H P 10, 000 6, 000 4, 000 H S+P Sma. I H+P 5, 000 12, 000 17, 000 1, 0 S S+H Uncut S 6, 000 P