ex11 - Exercise 11 Analysis 1. A restriction enzyme is an...

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Exercise 11 Analysis 1. A restriction enzyme is an enzyme that cuts through double stranded DNA at a  specific sequence in the DNA (restriction site). The DNA sequence that EcoRI  recognizes is (5’->3’) GAATTC and the complementary strand (3’->5’) CTTAAG. 2. 20 µL of plasmid DNA, 2.5 µL of 10x buffer and 2.5 µL of RNAse Final volume = (20+2.5+2.5 µL)=  25 µL C 1 V 1 =C 2 V 2           C = (V 1 /V 2 ) * C 1 Final concentration of 10x buffer = (volume of buffer/total volume)*concentration (2.5µL/25µL)*10=  1x 3. 18 µL of plasmid DNA, 2.5µL of 10x buffer, 2.5 µL of RNAse, 2 µL of EcoRI Final volume = (18+2.5+2.5+2 µL)=  25 µL Final concentration of 10x buffer= (2.5 µL/25 µL)*10=  1x 4. omit 5. Bacteria can be made competent by simple treatments of Ca++, followed by  incubation on ice with the ligated plasmid preparation, and finally being heat 
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This note was uploaded on 11/30/2008 for the course BIO 205L taught by Professor Hanson during the Fall '07 term at University of Texas at Austin.

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ex11 - Exercise 11 Analysis 1. A restriction enzyme is an...

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