BIOLOGICAL SCIENCES 231 Genomes and Genetic Analysis - 1 2 BIOLOGICAL SCIENCES 231 Genomes and Genetic Analysis Laboratory(Spring Semester 2014 Section

BIOLOGICAL SCIENCES 231 Genomes and Genetic Analysis - 1 2...

  • Smith College
  • BIO 230
  • Assignment
  • cylau
  • 98
  • 50% (2) 1 out of 2 people found this document helpful

This preview shows page 1 - 5 out of 98 pages.

1
2
BIOLOGICAL SCIENCES 231: Genomes and Genetic Analysis Laboratory (Spring Semester 2014) Section 1: Tuesday 1:00pm - 3:50pm Section 2: Wednesday 1:10pm – 4:00pm Section 3: Thursday 1:00pm - 3:50pm Laboratory: Ford Hall 120 Textbook: Genomes and Genetic Analysis Laboratory Manual Instructor (Sections 1 & 3): Instructor (Section 2): Mary McKitrick Graham Kent Office: Office: Sabin-Reed 216 Telephone: 212-865-5041 Office Ext: 3820 Office Hours: TBA Office Hours: TBA E-mail: [email protected] E-mail: [email protected] Course Outline Week Laboratory 1 Laboratory Introduction Laboratory Safety Choose Laboratory Partners 2 DNA and RNA I Grind mouse liver tissue in liquid nitrogen 3 DNA and RNA II Mouse genomic DNA purification by Column Chromatography 4 No lab!!! 5 DNA and RNA III Mouse liver RNA purification by column chromatography Quantification and quality analysis of DNA and RNA by Ultra violet (UV) spectrophotometry 6 DNA and RNA IV Gel 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 I Amplification of a mouse gene sequence using RT-PCR to clone into an expression vector. Week Laboratory 3
7 DNA and RNA VI Analysis of PCR and RT-PCR results by gel electrophoresis Set up sequencing reactions of RT-PCR and PCR products Cloning II PCR purification of gene RT-PCR product. Quantification of gene RT-PCR product by spectrophotometer. Restriction Enzyme digestion of gene RT-PCR product. 8 DNA and RNA VI Sequence purification of RT-PCR and PCR sequencing reactions Cloning III Purification 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. coli cells. Plate transformed E. coli cells onto agar plates. Quiz 1 9 Spring Break! 10 Cloning IV Pick transformed colonies and grow in liquid media. Colony PCR to check for recombinant vectors. 11 Cloning V Isolate 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) 12 DNA Sequencing and Bioinformatics I Analyze DNA sequence results of recombinant plasmid. DNA Barcoding of Plants and Animals Isolation of DNA from fish (sushi) PCR amplification of COT genes Cloning VI Start protein expression in E. coli cells (Done by Instructor). 13 Cloning VII and Protein Expression Purification of recombinant expressed protein by column chromatography. Quiz 2 14 Cloning VIII and Protein Expression Polyacrylamide gel electrophoresis of expressed mouse protein.

  • Left Quote Icon

    Student Picture

  • Left Quote Icon

    Student Picture

  • Left Quote Icon

    Student Picture