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Author: Lynne Prost
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Biochemistry 551: Biochemical Methods
Lab 1: Structural Analysis of HCAII using PyMOL
Lab 1: BACKGROUND
Lab 1: PRELAB
Lab 1: PROCEDURE Part 1
Lab 1: PROCEDURE Part 2
Lab 1: PROCEDURE Part 3
Lab 1: PROCEDURE Part 4
LAB 1 MINI REPORT
Lab 2: PCR Amplification of HCAII and pETblue2
Lab 2: BACKGROUND
Lab 2: PRELAB
Lab 2: PROCEDURE Part 1
Wet Lab Logistics and Policy Reminders
Lab 2: PROCEDURE Part 2
Lab 2: PROCEDURE Part 3
Lab 3: Analysis of PCR Products and Preparation for Gibson Assembly
Lab 3: BACKGROUND
Lab 3: PRELAB
Lab 3: PROCEDURE
LABS 2&3 MINI REPORT
Lab 4: Gibson assembly of HCAII into pETblue2 vector and transformation of E. coli with the reactions
Lab 4: BACKGROUND
Lab 4: PRELAB
Lab 4: PROCEDURE
Lab 5: Screening for pETblue2-HCAII clones
Lab 5: BACKGROUND
Lab 5: PRELAB
Lab 5: PROCEDURE
LABS 4&5 MINI REPORT
Lab 6: Induction of His-tagged HCAII expression
Lab 6: BACKGROUND
Lab 6: PRELAB
Lab 6: PROCEDURE
LAB 6 MINI REPORT
Lab 7: Purification of His-tagged wild type and mutant HCAII using Ni Resin
Lab 7: BACKGROUND
Lab 7: PRELAB
Lab 7: PROCEDURE
Lab 8: Analysis of His-tagged HCAII expression and purification
Lab 8: BACKGROUND
Lab 8: PRELAB
Lab 8: PROCEDURE
Lab 8: DATA ANALYSIS
LABS 7&8 MINI REPORT
Lab 9: Determination of Protein Stability using Chemical Denaturation and Intrinsic Fluorescence
Lab 9: BACKGROUND
Lab 9: PRELAB
Lab 9: PROCEDURE
Lab 9: DATA ANALYSIS
LAB 9 MINI REPORT
Lab 10: Investigation of the wild type and mutant HCAII enzyme activity
Lab 10: BACKGROUND
Lab 10: PRELAB
Lab 10: PROCEDURE
Lab 10: DATA ANALYSIS
LAB 10 MINI REPORT
Lab 11: FRET to detect ligand binding to HCAII
Lab 11: BACKGROUND
Lab 11: PRELAB
Lab 11: PROCOTOL
Lab 11: DATA ANALYSIS
Lab 11: FINAL LAB REPORT
Appendix 1: Information on HCAII
Appendix 2: Statistical methods used in 551 data analyses
Appendix 3: Additional laboratory details
Appendix 4: Buffer formulations
Appendix 5: Operation of the Shimadzu spectrophotometers
Lab 12: Independent Lab
Biochemistry 551 Lab Manual Copyright © by Lynne Prost. All Rights Reserved.
