Biotech

  1. Be able to create a standard curve using known fragment sizes (semi-log graph paper)!


  1. If given electrophoresis results, be able to approximate unknown plasmid sizes using the standard curve – you will be asked to us two different colors to interpolate fragment sizes just like we did in this lab. You will have to be within a range of acceptability to get points!


  1. Be able to figure out how many restriction sites there are on a plasmid or linear DNA given Gel results


  1. Be able to read a grid of reagents that are placed into microcentrifuge tubes and explain the purpose of each tube.


  1. Be able to explain the purpose of restriction buffers and incubation of DNA and restriction enzymes at 37°C for 45 minutes.


  1. Be able to explain the 3 purposes of TBE buffer in the lab


  1. Be able to explain why the proteins --- bromophenol blue and xylene cyanol move at different rates in a gel and provide the approximate sizes of equivalent DNA fragments in 0.9% agarose gels.


  1. Be able to explain the relationship of agarose percentage and separation of DNA fragments that are close in size


  1. Be able to explain the three purposes of loading dye – if you put down that it stains DNA, you will get an automatic zero on this test!


  1. Be able to explain the purpose of electricity in gel electrophoresis and the exact subunit of DNA that allows it to move with electricity.


  1. Be able to explain two purposes of restriction enzymes for bacteria in nature.


  1. Be able to explain how restriction enzymes are named and know the three restriction enzymes (Genus / species/ strain) that we used in the lab


  1. Know who coined the term restriction enzymes and why they are called this


  1. Be able to explain why smaller DNA fragments move at a faster rate than larger DNA fragments

  1. Be able to explain the difference between interpolation and extrapolation. Also, be able to explain why interpolation of unknown fragment sizes is more solid science!!


  1. Know what lambda is and the size of lambda DNA


  1. Know how prokaryotes prevent their own restriction enzymes from always cutting restriction sites in their own genome


  1. Be able to explain how scientists use sticky ends created by restriction enzymes to create recombinant DNA and recombinant organisms.


  1. Be able to explain the purpose of ethidium bromide in this lab. (micro and macro)


  1. Be able to explain why scientists electrophorese proteins, RNA, and DNA


  1. Be able to explain what RFLP stands for and explain how RFLP’s are used for genetic analysis


  1. Be able to look at gel results and create a plasmid map – (worksheet). Also be able to look at a plasmid map and show what the gel results would look like –(worksheet)


  1. Be able to use frequencies of alleles in a population to figure out the probability that a person has a specific allelic genotype using the rule of multiplication.


  1. Know ALL the questions in the restriction analysis lab and be able to apply that knowledge to new situations.