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Restriction Mapping.

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Presentation on theme: "Restriction Mapping."— Presentation transcript:

1 Restriction Mapping

2 Restriction Mapping: Double Digests
Factor I – Single Cut Factor 2 – Single Cut

3 Restriction Mapping: Double Digests

4 Restriction Mapping: Double Digests

5 Restriction Mapping: Double Digests

6 Single BamHI EcoRI HindIII PstI ScaI XbaI XhoI 5600 400 5200 600 1000 4000 1800 3800 100 900 1500 3100 2000 3600 2200 3400 500 1100 2400 3200 1600 1200 2800 2600 1400 1700 1900

7 Uses of Restriction Mapping
Fingerprinting Restriction Fragment Length Polymorphism Mapping

8 Cloning

9 pUC19

10 pUC19 Multiple Cloning Site

11 Ligation PLASMID TCGA AGCT KAN TCGA AGCT

12 Ligation DNA ligase PLASMID TCGA AGCT KAN TCGA AGCT

13 Ligation AGCT AGCT PLASMID KAN TCGA TCGA
WHY DO THIS AT LOW CONCENTRATION?

14 Ligation AGCT AGCT PLASMID KAN TCGA TCGA
WHY DO THIS AT LOW CONCENTRATION?

15 Ligation Reaction 1 Reaction 2 Reaction 3 pUC19 digest (50 ug/ul) 2 ul
RDK1984 digest (70 ug/ul) 6 ul 10X ligase buffer 1 ul water 7 ul T4 DNA ligase

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