POST LAB & ANALYSIS pGLO Bacterial Transformation & Gel Electrophoresis

Page 42: Lesson 2 Review Questions (do not include these questions in your Formal Lab 1. On which of the plates would you expect to find bacteria most like the original non-transformed E. coli colonies you initially observed? Explain your predictions 2. If there are any genetically transformed bacterial cells, on which plate(s) would they most likely be located? Explain your predictions. 3. Which plates should be compared to determine if any genetic transformation has occurred? Why? 4. What is meant by a control plate? What purpose does a control serve?

Page 43: Lesson 3 Data Collection & Analysis 1. Carefully observe and draw what you see on each of the four plates. Put your drawings in the data table below. A. Be sure to include: a)How much bacterial growth do you see on each plate, relatively speaking? b)What color are the bacteria? c)How many bacterial colonies are on each plate (count the spots you see) Colonies v Lawn

Page 44, 45 & 46: Analysis of Results Complete questions 1-4 on pg 44 Complete questions 1-4 on pg 45 Complete questions 1-3 on pg 46 Include these questions in your lab report!

Lab 2: Gel Electrophoresis

Warm-UP Today’s PLan What was your favorite part of the lab? What did you learn the most? Finish Lab….Please have out and ready, both labs! Start Inheritance Lesson Today

A B C D Restriction digest gels

A B C D Restriction digest gelsNote, these gels were not destained fully

Page 34: Analysis of Results In the first rectangle please trace a copy of your gel In the second rectangle please cut out ALL photocopies of the gels and tape in with your groups on top.  DO NOT GLUE YOU WILL NEED THIS FOR YOUR FORMAL LAB REPORT!

Page 36: Measurements from gel Measure from bottom of well to bottom of DNA Estimate size using HindIII…PUT answers in parenthesis in data chart…follow Mrs. Wait’s directions

Page 40: Semilog Graph Paper 1.Fragment size on Y. Distance traveled on X 2.Using the fragments from the lambda HindIII, plot the distance traveled in relationship to fragment size. This will be your standard curve which you will use to determine your unknown fragment size from the other three samples. Order on your gel picture is: L= LambdaE = EcoRI P = PstIH = HindIII

Page 36: Add Measurements from Semilog to your chart on pg 36 PUT answers from semilog in data chart on pg 36 under the parenthesis…follow Mrs. Wait’s directions (35,000) 34,500 14

Factors Affecting Restriction Enzyme Digestion Temperature, restriction enzymes are sensitive to prolonged periods of exposure to heat Cross contamination of restriction enzymes Buffer, optimum pH Incubation temperature, maintain optimum temperature during restriction enzyme activity

Pg 42: Evaluation of Results Complete the three questions on pg 42. Include these questions in your lab report!

Your Grade for this Lab! This will be 2 formal lab write ups. One for each lab. Both must be typed! See your INB formal lab write up More details It says you can write in pen…this is not true for this lab report! Include all questions as indicated in this power point. Rough drafts due Monday, May 16 th and Tuesday, May 17 th Final due Wednesday, May 25 th and Thursday, May 26 th. Must include data tables from gel electrophoresis Must include semi log graph Must include observations in data chart from lab 1