L6 - Lab 6 - GMB II - DNA Electrophoresis, Complementation...

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DNA Electrophoresis, Complementation and Genetics Readings: Lab Manual pgs 60-86 (81-84), 107-114 (112-114), 141-153. Campbell Chapters 12-17 (all but about 8 pages overlap with the 8-9 AM lecture) and Chapter 20.
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GMB II- outline DNA electrophoresis DNA charged, migration dependent upon size and conformation Visualize DNA bands Generate standard curve, determine size unknown Complementation- score growth. Mendelian Genetics Understand meiosis, homologous chromosomes. Alleles are positioned on chromosomes. Notation - unlinked traits versus linked traits. Making predictions: Punnett Squares.
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Lab Exam 1: Th 10/16 6:40 -8:20 Study Resources: EXAM READER 1) Previous lectures (webcast) 2) Reviews- Friday 10/3 8-10 PM, Friday 10/10 6-8 PM. Monday 10/13 5-6:30 PM. 3) Practice problems 4) Punnett squares 5) Webcast (Archives). Handout under study resources (lab exam 1).
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Office Hours (2084 VLSB) Mikes: Week of 10/6 office hours are M 11-1, W 1-2. Week of 10/13 office hours are M 11-1, T 2-4, W 1-3, Th 10-12. Lab GSIs: M 2-4, T 8: 30 -9: 30 , 1-2, 5: 30 -6: 30 , W 1-2, 5: 30 -6: 30 Th 8: 30 -9: 30 , 1-2, 5: 30 -6: 30 , F 1-2 Disc. GSIs: M-F 10-2 Study resources - webcasts, archived webcasts, reviews, office hours.
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0 10,000 20,000 30,000 40,000 48,502 LAMBDA Bam H1 SITES 5505 22,346 27,972 34,499 41,732 5505 16,841 5626 6527 7233 6770 21,226 21,226 26,104 31,747 39,168 44,972 4878 5643 7421 5804 3530 Eco R1 SITES Hind III SITES 23,130 25,157 27,479 36,895 37,459 37,584 44,141 23,130 2027 2322 9416 564 125 6557 4361 Restriction Maps of the Linear Genome ! ! ! maps: note bottom = Hind III 560 bp 2.0 Kbp 2.3 Kbp 4.4 Kbp 6.6 Kbp 9.4 Kbp 25 Kbp BamH 1 EcoR 1 HinD III * (25 Kbp band represents two pieces, 27.5 and 23.1 Kbp respectively that co-migrate). Purified pieces Amp R Isolate pBLU DNA Isolate source DNA R.E. digestion R.E. digestion
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+ pole Migration is dependent upon size & shape!
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0 10,000 20,000 30,000 40,000 48,502 LAMBDA Bam H1 SITES 5505 22,346 27,972 34,499 41,732 5505 16,841 5626 6527 7233 6770 21,226 21,226 26,104 31,747 39,168 44,972 4878 5643 7421 5804 3530 Eco R1 SITES Hind III SITES 23,130 25,157 27,479 36,895 37,459 37,584 44,141 23,130 2027 2322 9416 564 125 6557 4361 Restriction Maps of the Linear Genome ! ! No Enzyme Eco R1 Bam H1 Hin DIII Student pBLUi Pre-cut pBLUi Pre-cut pBLU Un -cut pBLU pBlui insert Hin DIII sites Diluted dye Diluted dye AAGCTT TTCGAA
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2 3 4 5 6 7 8 9 10 Dye ! - ! B ! E ! H Student pre-cut pre-cut un-cut dye pBLUi pBLUi pBLU pBLU Surface of Buffer (1X TBE) ! - ! B 1 2 3 4 5 6 7 8 9 10 ! E Figure 1: Loading the gel. Note the order of loading of the samples. Lanes 1 and 10 get diluted loading dye. Lanes 2-5 get various forms of ! DNA. Lanes diluted dye
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This note was uploaded on 09/28/2009 for the course BIO 1al taught by Professor Pederson during the Fall '08 term at University of California, Berkeley.

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L6 - Lab 6 - GMB II - DNA Electrophoresis, Complementation...

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