POST LAB & ANALYSIS pGLO Bacterial Transformation & Gel Electrophoresis.

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 44Complete questions 1-4 on pg 45Complete 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

AB

C D

5-2-2011 Restriction digest gels

AB

C D

5-3-2011 Restriction digest gels Note, 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 DNAEstimate size using HindIII…PUT answers in

parenthesis in data chart…follow Mrs. Wait’s directions

Page 40: Semilog Graph

Paper1. Fragment size on Y.

Distance traveled on X2. 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= Lambda E = EcoRIP = PstI H = 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)34,50014

Factors Affecting Restriction Enzyme Digestion

Temperature, restriction enzymes are sensitive to prolonged periods of exposure to heat

Cross contamination of restriction enzymes

Buffer, optimum pHIncubation 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 upMore 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 16th and Tuesday, May 17th

Final due Wednesday, May 25th and Thursday, May 26th. Must include data tables from gel electrophoresis Must include semi log graph Must include observations in data chart from lab 1