Download Gene Cloning

Lesson Plan Summary Sheet
Teacher:
Howerton
Subject:
Biotech 2
Topic:
Unit B2: Cloning
Date
Essential Question(s)
Create ppt slides for each LL/vocab
Activities
Vocabulary
Learning Logs
10/5
What are the major steps in cloning a PCR
amplified gene into E.coli
□ Overview the process of cloning a PCR
amplified gene
□ Look up information on PCR and create a
presentation on it
□ Pre-lab the PCR portion of the lab
PCR
Primer
Template
Taq polymerase
Thermocycler
Denaturation
Annealing
Degenerate primers
Nesting primers
RT-PCR
qPCR
C21: Create a presentation
that uses the vocabulary to
describe the technique and
uses of PCR
10/6
How do you amplify a target DNA using the
polymerase chain reaction technique?
□ Amplification of a 270bp Target DNA
Fragment by PCR
□ Look up information on gel electrophoresis
and create a presentation on it
□ Pre-lab the gel analysis portion of the lab
C22: Create a presentation
that uses the vocabulary to
describe the technique and
uses of agarose gel
electrophoresis.
10.7
How do you use gel electrophoresis to analyze a
PCR product?
□ Analysis of PCR product by agarose gel
electrophoresis
□ Look up information on restriction digests
and create a presentation on it
□ Pre-lab the clean and cut portion of the lab
Agarose
Electrophoresis
Buffer
Combs
Wells
Standards
Percentage
Loading dye
Stain
Restriction enzymes
Palindromes
Optimum
Cleavage
Restriction site
Sticky end
Blunt end
C23: Create a presentation
that uses the vocabulary to
describe the technique and
uses of restriction digest
analysis.
10/8
How can spin columns and restriction digest be
used to prepare the PCR amplified insert for
ligation?
□ Preparation of insert for ligation (spin
column cleaning and restriction digestion)
□ Look up information on DNA ligation and
create a presentation on it
□ Pre-lab the ligation portion of the lab
10/9
How are PCR amplified inserts ligated into a
vector plasmid?
□ Ligation of the PCR amplified 270bp DNA
insert into pUC19
□ Look up information on bacterial growth and
create a presentation on it
□ Pre-lab media preparation portion of the lab
10/12
How is media made up for transformation and
cell culturing?
□ Make agar plates and overnight broth tubes
for transformation
□ Look up information on transformations and
create a presentation on it
□ Pre-lab transformation portion of the lab
10/13
How are bacterial cells transformed with
recombinant DNA?
□ Transformation of ligated product into E.
coli cells
□ Look up information on culturing bacteria
and create a presentation on it
□ Pre-lab cell culturing portion of the lab
10/14
How can cells be grownup overnight to study?
□ Observe and calculate transformation
efficiency
□ Start overnight cultures
□ Look up information on plasmid isolation
Transgene
Insert
Vector
GMOs
Ligation
T4 DNA Ligase
Plasmids
Multiple cloning
region
Bidirectional insert
Unidirectional insert
Recombinant
Media
Agar
Broth
Sterile
Aseptic
Autoclave
C24: Create a presentation
that uses the vocabulary to
describe the technique and
uses of ligation
Transformation
Competent cells
Transfection
Transduction
Heat shock
Electroporation
Inoculate
Antibiotic screening
Efficiency
Selection
Culture
Exponential growth
Stationary growth
Scale-up
Lag phase
Log phase
Doubling time
Alkaline
Lysis
Neutralization
Precipitation
C26: Create a presentation
that uses the vocabulary to
describe the techniques and
uses of inserting DNA into
a cell.
C25: Create a presentation
that uses the vocabulary to
describe the technique and
uses of media preparation
C27: Create a presentation
that uses the vocabulary to
describe the techniques and
uses of culturing bacterial
cells.
C28: Create a presentation
that uses the vocabulary to
describe the techniques and
uses of plasmid isolation.
and create a presentation on it
□ Pre-lab plasmid isolation portion of the lab
10/15
How can plasmids be isolated from E.coli cells
for analysis?
□ Isolation of plasmids from transformed and
control cells
□ Pre-lab restriction analysis portion of the lab
No new terms
10/1619
How can plasmids be analyzed using restriction
digests?
No new terms
10/20
How do you determine the sequence of DNA?
□ Restriction analysis of recombinant plasmids
□ Look up information on DNA Sequencing
and create a presentation on it
□
□ Students will complete a simulation on the
Sanger method of sequencing
10/21
How genes be linked to diseases in a lab?
□
Lab: DNA Chips to Disease (Microarray)
kit
10/22
How do you analyze a cloned gene using
bioinformatics?
□
10/22
What methods are used to analyze RNA?
□
10/22
What techniques are used to manipulate DNA in
a biotech lab?
□
□
10/23
What techniques are used to manipulate DNA in
a biotech lab?
□
□
□
Students will complete the Bioinformatics
lab by using the BLAST to analyze the
sequence of their cloned gene.
Students will use internet to research
various RNA techniques
Review
Turn in Research Project Question,
Hyopthesis and Procedure
Turn in Lab Notebook
Test
Turn in Learning Log/Voca powerpoint
presentation
Sanger sequencing
Deoxynucleotides
Dideoxynucleotides
Microarrays
Bioinformatics
BLAST
See sheet
C29: Create a presentation
that describes the why and
how you can vary the
addition of the new insert
into the vector plasmids.
C210: Create a presentation
that uses the vocabulary to
describe the techniques and
uses of DNA sequencing
C211: Explain the Sanger
method of sequencing
C212: Explain the use of
DNA Chips (microarrays)
in research.
C213: Explain what BLAST
does.
C214: Explain three RNA
techniques
C215: Explain how you plan
to study for this test and
evaluate if you were
successful.
C216: Reflect on your effort
and what you have learned
in this unit.