50%(2)1 out of 2 people found this document helpful
This preview shows page 1 - 5 out of 98 pages.
BIOLOGICAL SCIENCES 231:Genomes and Genetic Analysis Laboratory(Spring Semester 2014)Section 1: Tuesday 1:00pm - 3:50pmSection 2: Wednesday 1:10pm – 4:00pmSection 3: Thursday 1:00pm - 3:50pm Laboratory:Ford Hall 120Textbook:Genomes and Genetic Analysis Laboratory ManualInstructor (Sections 1 & 3): Instructor (Section 2):Mary McKitrick Graham KentOffice:Office:Sabin-Reed 216Telephone: 212-865-5041 Office Ext: 3820Office Hours:TBAOffice Hours: TBAE-mail:[email protected]E-mail: [email protected]Course OutlineWeekLaboratory1Laboratory IntroductionLaboratory SafetyChoose Laboratory Partners2DNA and RNA IGrind mouse liver tissue in liquid nitrogen3DNA and RNA IIMouse genomic DNA purification by Column Chromatography4No lab!!!5DNA and RNA III Mouse liver RNA purification by column chromatography Quantification and quality analysis of DNA and RNA by Ultra violet (UV) spectrophotometry 6DNA and RNA IVGel electrophoresis of DNA and RNA Gene expression analysis of a mouse gene (transthyretin) using Reverse Transcription PCR (RT-PCR) Polymerase Chain Reaction (PCR) amplification of a mouse gene (transthyretin) from genomic DNA Cloning IAmplification of a mouse gene sequence using RT-PCR to clone into an expression vector.WeekLaboratory3
7DNA and RNA VIAnalysis of PCR and RT-PCR results by gel electrophoresisSet up sequencing reactions of RT-PCR and PCR products Cloning IIPCR purification of gene RT-PCR product.Quantification of gene RT-PCR product by spectrophotometer.Restriction Enzyme digestion of gene RT-PCR product. 8DNA and RNA VISequence purification of RT-PCR and PCR sequencing reactionsCloning IIIPurification of Restriction Enzyme digestions by organic extraction.Column purification of Restriction Enzyme digestions for size selection.Quantification of purified gene RT-PCR product and purified vector by spectrophotometer.Ligation of purified gene RT-PCR product to expression vector.Transform ligated mouse gene/vector to E. colicells.Plate transformed E. colicells onto agar plates.Quiz 19Spring Break!10Cloning IV Pick transformed colonies and grow in liquid media.Colony PCR to check for recombinant vectors.11Cloning VIsolate pure recombinant mouse gene/vector (plasmid prep).Quantify pure recombinant plasmid.Set up DNA sequencing reactions with recombinant plasmid.Purification of sequencing reactions (next day) 12DNA Sequencing and BioinformaticsIAnalyze DNA sequence results of recombinant plasmid.DNA Barcoding of Plants and AnimalsIsolation of DNA from fish (sushi)PCR amplification of COT genes Cloning VIStart protein expression in E. colicells (Done by Instructor).13Cloning VII and Protein ExpressionPurification of recombinant expressed protein by column chromatography.Quiz 214Cloning VIII and Protein ExpressionPolyacrylamide gel electrophoresis of expressed mouse protein.