Biochemical Protocol.8. DNA (2)

Genomic DNA preparation (plants)
Hybridization

Large Scale Plasmid Preparation: MGH

Modifications of DNA ends

PCR amplification of DNA

PCR product purification (Qiagen)

Perfectly Blunt Cloning of DNA (Novagen)

Phenol/chloroform extraction of DNA

Plasmid midi-prep from bacteria

Preparation of genomic DNA from bacteria

Purification of DNA

Q-PCR

Random Primed labeling of DNA (Prime-It II -Stratagene)

Restriction enzyme digestion of DNA
Southern Hybridization
Stark's Prehybridization Buffer

Sequencing using Sequenase

Southern blot

Tail DNA prep

Wizard Plus Mini-preps (109. Plasmid Mini-prep)

Worm Genomic DNA: MGH

Worm genomic Southern blots: Michael Koelle

In vivo transcription

BrUTP labeling of RNA transcripts

Biochemical Protocol.9. PCR (1)

About PCR

AFLP PCR

Alu-PCR

Asymmetric PCR

Colony PCR Protocols

Competitive and/or Quantitative RT-PCR

Degenerate PCR

Differential Display PCR

Genotyping by PCR

History of Polymerase chain reaction (PCR)--(1)

History of Polymerase chain reaction (PCR)--(2)

Important Publications at Cell

Important Publications at Nature

Important Publications at Science

In Situ PCR

Introduction to PCR

Inverse PCR

Ligation Mediated Suppression PCR

Long PCR Protocol: Church Lab

Long PCR Protocols

Methylation Specific PCR

Multiplex PCR

Nested PCR

PCR Animations

PCR aplications

PCR Brochure and Articles

PCR Elisa

PCR Manual

PCR Primer Design Tools

Biochemical Protocol.9. PCR (2)

PCR RFLP

PCR SSCP

PCR Troubleshooting

PCR, RT-PCR, and Real Time PCR Tutorials

Polymerase Chain Reaction (Molecular Info)

Polymerase chain reaction--PCR

RACE PCR

RAPD PCR

REAL TIME PCR Papers

Real Time PCR Protocols

Regular PCR Procedure

REP-PCR

RT PCR Protocols

Special PCR Protocols

Tail PCR

Tc1 Deletion PCR Tips: MGH

Touchdown PCR

Variants of PCR (1)

Variants of PCR (2)

Variants of PCR (3)

Variants of PCR (4)

Variants of PCR (5)

Vectorette PCR

Videos and Animations for PCR

Web Source of PCR (1)

Web Source of PCR (2)

Web Source of PCR (3)

Web Source of PCR (3)

Biochemical Protocol.10. Cellular Biology

Aseptic Cell Transfers

Calcium Phosphate tranfection of adherent mammalian cells (works well for 293 cells)

Cell mass by Measurement of Turbidity

Computation of Transfer Aliquots

Culturing MEFs
Calcium Phosphate Coprecipitation

Determining cell counts

Effectene (QIAGEN) tranfection of adherent mammalian cells (works well for HeLas and many other cell types)

Establishing stable cell lines

Establishment of a Primary Culture

Eukaryote Growth Dynamics

Examination of Bacterial Colonies

Fugene (Roche) tranfection of adherent mammalian cellsDOTAP (Roche) tranfection of adherent mammalian cells

Fusing cells to create heterokaryons

Gram Stain (+\-)

Harvesting murine bone marrow

Hematopoietic CFC assays

Maintenance

Metaphase chromosome preparation

Preservation and storage

Prokaryote Cell Number by Dilution Plating

Purification of L-KS+ marrow cells
Retrovirus Production
STO and ES Cell Culture

Safety considerations

Tissue culture methods

Tissue Culture of PtK1 Cells

Transfer of Eukaryote Suspension Cultures

TUNEL Protocol

Types of cells grown in culture

Viability Cell Count

Work area and equipment
XC assay of MoMLV

Biochemical Protocol.11. Cellular Organelle

MT/Organelle Motility Assay*
Membrane Mixes: Variations on a Theme*
Preparation of Rat Liver Cell Cytosol and/or Organelle Fractions*
Preparation of Sea Urchin Sperm Axonemes*
Preparation of Porcine Brain Tubulin*

Biochemical Protocol.12. Immunology

Antibody cleanup Yale Genome Analysis Center, Yale University

Bioengineering and Immunology (Video Protocol)

Computer immunology

Conjugation of monoclonal antibodies (complete) Dr. M Roederer, NIH, Vaccine Research Center

Direct Immunofluorescence Staining of Mononuclear Cells BD Biosciences

Direct Immunofluorescence Staining of Whole Blood Using Ammonium Chloride Lysing Solution BD Biosciences

Iindirect immunoflourescence for yeast I Dr. DE Koshland, Carnegie Institution of Washington

Immunofluorescence Protocols and Solutions

Immunofluorescence staining of cell suspensions of murine lymphoid tissue EBioscience.com

Immunofluorescence staining of human peripheral blood EBioscience.com

Immunofluorescence Technique Dr. DL Spector, Cold Spring Harbor Laboratory

Immunofluorescent Staining of Mouse and Rat Leucocytes BD Biosciences

Immunohistochemistry on Cryosections

Immunology Field Trip--Protocol

In Situ Probe

Indirect Immunofluorescence Staining of Mononuclear Cells BD Biosciences

Isolation of Resting B Cells From Mouse Spleen

MACS Depletion Drs. L Herzenberg, Stanford University

Making FITC and Biotin conjugates Therapeutic Immunology Group, Oxford University

Preparing cells for immunofluorescence Yale Genome Analysis Center, Yale University

Biochemical Protocol.13. Flow Cytometry Protocols (1)

7-AAD Staining of Dead Cells

A flow cytometric protocol for detection of Cryptosporidium spp.

Blocking FC-Receptor Binding

Calibrating Becton Dickinson Flow Cytometers for RFI Measurements

CD38 Fluorescence Intensity Quantitation

Detection of Apoptosis by 7-AAD Staining

DNA Staining for Cell Cycle Analysis

FCM: DNA Analysis by Flow Cytometry

FCM: FASC Analysis of Cell Cycle using BrdU and PI

FCM: Immunofluorescent Staining of Rat Thymocytes

FCM: Sodium and Potassium Analysis by Flow Cytometry

Flow Cytometry Glossary

Flow cytometry of murine blood

Flow Cytometry Principles （UC Berkeley）

Flow Cytometry Protocol

Flow Cytometry Protocol for Intracellular Staining Using Conjugated Second Antibody.

Flow Cytometry Protocol for Staining of Colonocytes - Surface Antigens

Flow Cytometry: Calcium Flux Analysis

Flow Cytometry: Membrane Potential Analysis by Flow Cytometry

Flow Cytometry：Annexin V Protocol

Flow Cytometry：CaspaTag Caspase Activity Protocol

Flow Cytometry：PARP PE

GENERAL CELL STAINING PROTOCOL FOR FLOW CYTOMETRY 1)