Transcript pGLO Bacterial Transformation & Gel Electrophoresis

pGLO Bacterial Transformation
& Gel Electrophoresis
POST LAB & ANALYSIS
Page 42: Lesson 2 Review Questions (do not
include these questions in your Formal Lab
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?
1.
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)
b)
c)
How much bacterial
growth do you see on
each plate, relatively
speaking?
What color are the
bacteria?
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
Lab 2: Gel Electrophoresis
Today
Warm-UP
Today’s PLan
 What was your favorite
 Finish Lab….Please
part of the lab?
 What did you learn the
most?
have out and ready,
both labs!
 Start Inheritance
Lesson
5-2-2011 Restriction digest gels
A
C
B
D
5-3-2011 Restriction digest gels
A
C
Note, these gels were not destained fully
B
D
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
Order on your gel picture is:
L= Lambda
E = EcoRI
P = PstI
H = HindIII
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.
Page 36: Add Measurements from Semilog to
your chart on pg 36
14
(35,000)
34,500
 PUT answers from semilog in data chart on pg 36
under the parenthesis…follow Mrs. Wait’s directions
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.
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 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
