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.