campbl20 - Overview Restriction Endonucleases A Restriction...

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Overview
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Restriction Endonucleases A Restriction Endonucleases will cut both strands of a DNA duplex at a specific place These “places” need not be directly opposite: Note that the above enzyme is Eco RI, the first restriction endonuclease characterized 5’…GAATTC…3’ 3’…CTTAAG…5’ 5’…GAATTC…3’ 3’…CTTAAG…5’ 5’…G -OH P- AATTC…3’ 3’…CTTAA -P HO- G…5’
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Sticky Ends
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Ligation
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More RE Enzymes Enzyme Sequence Product Eco RI G^AATTC 5’ sticky ends Bam HI G^GATCC 5’ sticky ends Bg1 II A^GATCT 5’ sticky ends Pvu I CGATC^G 3’ sticky ends Pvu II CAG^CTG Blunt end Mbo I G^ATC 5’ sticky ends Hind III A^AGCTT 5’ sticky ends Hinf I G^ANTC 5’ sticky ends Sau 3A G^ATC 5’ sticky ends Alu I AG^CT Blunt end Taq I T^CGA 5’ sticky ends Hae III G^GCC 5’ sticky ends Nof I GC^GGCCGC 5’ sticky ends
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Most R.E. Recognition Sequences are Palindromes G^AATT-C C-TTAA^G G^GATC-C C-CTAG^G A^GATC-C T-CTAG^G GC^GGCC-GC CG-CCGG^CG Nodeba Bob Abedon
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Ligation into Plasmid Upon Ligation, we now have Recombinant DNA !
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campbl20 - Overview Restriction Endonucleases A Restriction...

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