MOLECULAR BIOLOGY LABORATORY MANUAL

Link: http://www.aecom.yu.edu/cellbiology/BCL6/BHY%20Lab%20Manual.doc

A COMPILATION OF METHODS CURRENTLY USED IN THE LABORATORY OF
DR. B. HILDA YE

TABLE OF CONTENTS

1. Oligolabeling (Random priming)
2. Screening Recombinant Phage Libraries
3. Total RNA Preparation
4. Phage Mini-prep
5. PEG Plasmid Prep
6. Extraction of DNA from Low Melting Point Agarose Gels
7. Plasmid Cloning: Ligation and Transformation
8. Preparation of Plasmid Cloning Vectors
9. Preparation and Freezing of Competent Bacteria
10. Lambda Phage Prep
11. Screening Library with Oligonucleotides Probes
12. Preparation of Polyacrylamide-Urea "Sequencing" Gels
13. Southern Blotting and striping filters
14. DNA Sequencing
15.Field Inversion Gel Electrophoresis
16.DNA Molecular Weight Markers
17.Retroviral Gene Transfer: Infections Cells in Suspension
18.Titration of Virus-producing Cells
19.Run-Off Transcription
20. RNase Protection Assay
21.Northern Blotting
22.Freezing Bacteria
23.Multi-gene Expression Assay (Reverse Northern Slot Blot)24.Mini-prep Plasmid DNA Protocol for Sequenase
25.Purification of Crude Oligonucleotides
26.Preparation polyA+ mRNA
27.Direct sequencing of PCR Amplified Fragments
28.Oligomer Purification for Trityl off Synthesis
29.Protocol for Race-PCR
30.Electroporation and selection of human B and T cells
31.CAT assay
32.Beta galactosidase assay
33.Transfection by Ca-P Precipitation
34.Agar cloning
35.Freezing tissue cultured cells
36.Titration of virus producing cells (simplified method)
37.Titration of virus producing clones by RT-PCR
38. Recipes of Cell Culture
39.Pulse Field Gel Electrophoresis
40.SSCP (Single Strand Conformation Polymorphism
41. Immunofluorescence (HA)
42.Aldh Assay
43.Instructions for Blotter (Genie)
44.Suggestions on How to scan a Picture
45. Protocol for SDS-PAGE (Laemmli)
46.Cisplatin Resistance Assay
47.MTT Assay
48. 96 well microplate staining for FACS analysis
49. 96 well microplate nuclear staining for FACS analysis
50. Gel mobility shift assay (EMSA)-for BCL-6
51.Rapid mouse tail tip DNA extraction protocol
52.Preparation of total DNA from apoptotic cells
53.Starvation of Rat-1 myc cells
54. Stain for Beta-Galactosidase Expression in Cell Culture
55. Procedures for Inducing Gene Expression from MT Promoter