Experiments of Biochemistry and Molecular Biology

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

  1. Molecular Cloning: A laboratory Manual, 4TH Edition, Cold Spring Harbor Press
  2. Short Protocols in Molecular BiologyEdited by Frederick M. Ausubel, Roger Brent, Robert E. Kingston, David D. Moore, J.G. Seidman, John A. Smith, and Kevin Struhl 

Course Website