Laboratory Manual for Biomedical Research

First Edition

Chapter 1. General Lab Techniques

Lab security and basic techniques

Advanced lab skills

Advanced lab skills (2)

Chapter 2. Molecular Biology -- Molecular Separation & Labeling

Molecular Separation

• DNA Agarose Gel Electrophoresis 
• RNA Agarose Gel Electrophoresis 
• SDS-Page - Polyacrylamide gel electrophoresis 
• 2D Page-Two - Dimensional Polyacrylamide Gel Electrophoresis 
• Ion Exchange Chromatography 
• Gel Filtration Chromatography 
• Affinity chromatography 

Chapter 3. Molecular Biology -- DNA and RNA

Nucleic acid methods (1)

Nucleic acid methods (2)

DNA isolation & related protocols

DNA Purification (glass milk vs electroelution)

DNA, RNA Sequencing

Recombinant DNA Techniques

RNA Isolation and Purification

Isolation of DNA,RNA, and Protein simultaneously.

DNA mutation detection by SSCP

Preparation of DNA and RNA probes

Southern blot hybridization

Northern blot hybridization

Loss of Heterozygosity (LOH)

Gene knockout protocol

SiRNA gene knockout

Plasmid and its usefulness

DNA library construction

Microarray protocols.

• Basic knowledge of microarray.
• Introduction to Microarray.
• MicroArray Procedure
• Total RNA Isolation from cultured cells.
• DNase Treatment of Total RNA
• Making the single strand cDNA probe.
• Automated Slide Processor (ASP) Version for hybridization.
• Washing microarrays in ASP.
• Processing of Array slide
• Pre-hybridization of the processed slides (NON-Automated version).
• Hybridization of Cy3 + Cy5 probe to glass array (NON-Automated version).
• Preparation of Dendrimer Cy3 and Cy5.
• Washing unbound probe from glass array (NON-Automated version).
• Hybridization of Dendrimers (Cy3 and Cy5) to Array (NON-Automated version).
• Washing unbound dendrimer from glass array(NON-Automated version).
• Microarray Dababases
• Troubleshooting
• Other Microarray Protocols (1,2)

Gene transfection

Gene therapy for cancer

Molecular cloning

Conditional gene transfection(Tet on/off)

Chapter 4. Genetics

Epigenetics protocols

Mutagenesis protocols

Single nucleotide polymorphisms (SNPs)

Chapter 5. Molecular Biology --PCR Serials

PCR,RT-PCR,Real time PCR etc.

PCR (General Procedure)

• Recommended Reagent Concentrations
• Recommended Reaction Conditions
• Initial Conditions
• Temperature Cycling
• "Hot Start" PCR
• Asymmetric PCR for ssDNA Production
• Detecting Products
• Labelling PCR Products with Digoxigenin
• Cleaning PCR Products
• Sequencing PCR Products
• Cloning PCR Products
• AND ALWAYS REMEMBER

PCR Primer Design Tools

RT-PCR

Real time PCR

More PCR Protocols Online

Video and Animation of PCR

Mouse Genotyping by PCR

PCR Based Molecular Cloning

Variants of PCR

PCR Troubleshootings

Chapter 6. Molecular Biology -- Protein

Protein methods

Enzyme Assay Protocols(1)   (2)   (3)

Protein labeling techniques

Protein sequencing

Dot Blot

Subcellular fractionations

Western blot hybridization

• Western Blot Hybridization Center
• Western Blot Full Procedure (from Pierce)
• Western Blotting Procedure with Video Show
• Protocol 1: Western Blot
• Sample protein preparation
• Protein concentration assay
• Electrophoresis and blotting
• Blocking non-specific antigen
• Incubation with primary antibody
• Incubation with secondary antibody
• Substrate (ECL) incubation
• Restore a western blot using pierce stripping buffer
• Western blot related solutions and buffers
• Western blot handbook
• Western blot troubleshooting
• Protocol 2: Western Blot 2.
• Protocol 3: Western Blot 3.
• More online Western blot protocols (1) (2)

Protein chips

Kinase assay

Methods for detecting protein phosphorylation

Chapter 7. DNA Protein Interactions

EMSA

ChIP assay