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Lecture 14

CAS BI 216 Lecture 14: Genetics - Lecture 14 - Chapter 9
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Department
Biology
Course
CAS BI 216
Professor
John Celenza
Semester
Spring

Description
Lecture 14 33017 (Chapter 9) Chapter 9 Recombinant DNA Restriction Endonucleases (R.E.) Sticky 5 Overhang 5 NNNN 3 5 NNNNN 3 3 NNNNNNN 5 3 NN 5 Sticky 3 Overhang 5 NNNNNNN 3 5 NNN 3 3 NNNN 5 3 NNNNNN 5 Blunt Ends 5 NNNN 3 5 NNNN 3 3 NNNN 5 3 NNNN 5 o R.E. recognize unique sequences EcoRI 5 GAATTC 3 3CTTAAG 5 o ~200 R.E. one can purchase Average Fragment size o probability find restriction site (14)^n (n= bases in recognition site) o examples EcoRI GAATCCTTAAG probability (14^6) > 14096 average fragment size = 4096 bp ~ 4000 bp 4 Bases (14 ^ 4 > 1256 average 256 bp fragment sizes ~ 250 bp 8 bases 14 ^ 8 > 165,536 average fragment size = 65,536 bp ~ 50,000 Detection using gel electrophoresis o Ethidium Bromid binds tightly to DNA (EtBr) in gels EtBr detects minimally ~1ng of DNA (1 ng = 1 nanogram = 10 grams) Example 1000 bp dsDNA fragment o AT and GC base pairs have molecular weight (660 Daltons) 1 1 1 660 1000 10
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