Course Name: Experiments of Biochemistry and Molecular Biology
Credits: 3.0
Level: Undergraduate
Pre-requisite: Organic Chemistry, Biochemistry, Molecular Biology
Lecture Time: 12 weeks, 1 session/week, 5 hours/session
Instructors: Dr. Guohong Li
Course Description
This course introduces basic principle and experimental techniques in Biochemistry and Molecular Biology, including protein purification and identification, enzyme kinetics, molecular cloning and RNA techniques etc. This course is one of the obligatory major courses for undergraduate students majoring Life Sciences, and is also one of the elective courses for other disciplines, such as Chemistry and Physics.
Topics and Schedule
Experiment 1. Protein determination and measurement of protein concentration
(5 hrs)
SDS-PAGE, Coomassie blue staining, Western Blots
Experiment 2.Identification and purification of recombinant proteins by His6- or GST- affinity chromatography (5 hrs)
affinity chromatography, SDS-PAGE
Experiment 3.Identification and purification of proteins by molecular sieve (5 hrs)
ÄKTA start system for protein purification
Experiment 4.Determination of the activity and kinetic constants of catalase (5 hrs)
enzyme kinetics
Experiment 5.Determination and identification of nucleic acid concentration (5 hrs)
UV spectrophotometric determination, agarose gel electrophoresis
Experiment 6.Extraction and identification of plasmid DNA (5 hrs)
Plasmid extraction, agarose gel electrophoresis
Experiment 7.Restriction enzyme digestion and identification (5 hrs)
Restriction enzyme digestion, agarose gel electrophoresis
Experiment 8.Purification, isolation and ligation of plasmid DNA restriction fragment (5 hrs)
ligation of DNA, agarose gel electrophoresis
Experiment 9.Preparation of competent cells and transformation of ligated plasmid DNA (5 hrs)
Competent cells, transformation
Experiment 10.Screening of the transformation by PCR (5 hrs)
PCR
Experiment 11.Extraction and identification of RNA by agarose gel electrophoresis
(5 hrs)
RNA extraction
Experiment 12.Reverse transcription polymerase chain reaction (RT-PCR) (5 hrs)
RT-PCR
Grading
The performance of experimental operation in each class will be evaluated and graded, their scores will count for 40% of the total. At the end of the course will be followed by a final examination of experimental operation, which will count for 60%.The final scores are graded by excellent, good, qualified and failed.
Textbook
Self compiled lecture notes by instructors
References
Course Website
