included Protocols

Category: Plant Biology Protocols: Arabidopsis Tissue Culture

Experiments included for analysis of plant epidermis: Steroid induction of gene expression in Arabidopsis; Replica Molds and Casts of the plant epidermis. - [Read Analysis of the Plant Epidermis Experiments]

Category: Chromatography Protocols: Column Chromatography Protocols

Column Chromatography Protocol DOC, includes troubleshooting information. Equipment used included: Peristaltic pump, UV detector , Chart recorder, and Column. Prestegard Lab. - [Read Column Chromatography Protocol DOC]

Category: Epigenetics and Methylation Protocols: Bisulfite Treatment Methylation Assay Protocol

A detailed protocol for bisulfite treatment of DNA. DNA is first digested with a restriction enzyme. DNA is denauted at 97C for 5min. Bisulfite protocol is included. Also DNA purification using the Promega Wizard DNA Cleanup kit in indicated. Fan Lab. UCL - [Read Fan Lab Bisulfite Treatment of Genomic DNA]

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This protocol describes the first step in constructing an array: amplification of the predicted ORFs that are to be included in the array. Gene-specific primers containing vector-specific flanking sequences that facilitate recombinational cloning are used to amplify each ORF. A secondary amplification can be used to extend the length of the homologous vector sequence flanking the ORF. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: Amplification of ORFs]

Category: Chromatography Protocols: Liquid Chromatography Protocols

Protocols and information on general HPLC column care. Includes: Silica based columns and Polymer based columns.  Protocols included: pH stability, Mechanical stability, Mobile phases (Eluents), Proper storage of HPLC columns, Equilibration time, Regeneration of a column, Regeneration of RP packings, Regeneration of NP (Normal Phase) packings, Regeneration of Ion Exchange Packings - [Read HPLC Column Care Protocols and Information]

Category: Cell Biology and Cell Culture: ES Stem Cell Biology and Culture

hES - Human Embryonic Stem Cell Culture Manual. Excellent Manual with Pictures. Troubleshooting guide included. Many Topics Included. ES Cell International Pte Ltd. - [Read Human Embryonic Stem Cell Culture Manual]

Category: Immunology: T Cell Protocols

Describes methods for labeling high or low numbers of lymphocytes with CSFE. Protocols are provided to use CSFE-labeled cells in cell transfer studies or as cells to be cultured in vitro. Detailed guidelines for positioning of CSFE-labeled lymphocytes in lymphoid organs or other tissues are included for those wishing to use this approach to study lymphocyte migration. - [Read Intracellular Fluorescent Dye CFSE to Monitor Lymphocyte Migration and Proliferation]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Retroelements and their derivatives are a ubiquitous and abundant component of plant genomes. Major classes of retroelements include the Pseudoviridae (Ty1-copia ), the Metaviridae (Ty3 -gypsy) and the Retroposineae LINE (non-LTR) groups. All reverse transcribing elements can be included in a universal classification. Includes: Pseudoviridae (Ty1-copia) Degenerate Primers; Metaviridae (Ty3-gypsy) Element Degenerate Primers; LINE Element Degenerate Primers; PCR Programmes. - [Read Isolation of Retroelement from Plant Genomic DNA]

Category: Neuroscience Protocols

Protocols included: USE OF RADIAL ARM MAZE TASK TO TEST BASIC WORKING MEMORY; USE OF RADIAL ARM MAZE TASK TO TEST WORKING VERSUS REFERENCE MEMORY; USE OF MORRIS WATER MAZE TASK TO TEST SPATIAL MEMORY; USE OF WATER MAZE TASK FOR SPATIAL PROBE TRIAL; USE OF WATER MAZE TASK TO TEST WORKING MEMORY. - [Read Learning and Memory Protocols]

Category: C. elegans Protocols

To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. - [Read Live Imaging of Caenorhabditis elegans: Examples]

Category: Yeast Cell Culture Protocols: Yeast Media, Solutions and Stocks Protocols

The yeast, Saccharyomyces cerevisiae, has become an important organism in molecular, biochemical, and genetic analysis. The organism has specific requirements for growth under a variety of conditions. The media, both liquid and solid, simple, define, and complex are describe in this unit. Also included are methods for handling, storing, and shipping stock of yeast. - [Read Media and Culture of Yeast Protocol]

Category: DNA Protocols: EMSA DNA-Protein Protocols

Nuclear Extract Preparation for EMSA after Wadman et al., EMBO J. 1997. Buffer recipies included. - [Read Nuclear Extract Preparation for EMSA (PDF)]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols

A procedure for direct and indirect staining of single-cell suspensions of lymphoid tissue or peripheral blood lymphocytes to detect cell surface membrane antigens is presented. In addition, support protocols present methods for fluorescence labeling of purified antibodies. A protocol for flow cytometric analysis of intracellular antigens in single-cell suspensions is also included. - [Read Preparation of Cells and Reagents for Flow Cytometry Protocols]

Category: Signalling Signal Transduction Protocols

The protocol provides a method to achieve a sample sufficient for one determination of cAMP using the acetylation protocol. The protocol describes the method used for measuring the content of cyclic adenosine 3',5'- monophosphate (cyclic AMP or cAMP) in RAW 264.7 cells using an enzyme-linked immunoassay system. Information included in the protocol: Treatment of Cells and Preparation of Extracts; Reagents and Materials. - [Read Preparation of RAW 264.7 Lysates for Cyclic AMP Determination]

Category: Microscopy Protocols: In Vivo Imaging Protocols

To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. Chromatin dynamics can be followed easily, and wild-type embryonic cells can be compared with mutants or RNAi-treated embryos. - [Read Protocol Live Imaging of Caenorhabditis Elegans]

Category: Signalling Signal Transduction Protocols: MAPK Protocols

This protocol uses mitogen-activated protein/ERK kinase (MEK) to activate the extracellular-signal-regulated (Erk) mitogen-activated protein (MAP) kinases upon agonist binding to receptors. Protocol includes information about: Harvest, Immunoprecipitation and Kinase Reaction. Also includes following solutions: Laemmli Sample Buffer (1X), ATP Mix, Kinase Buffer (10X), Lysis Buffer. Helpful protocol hints are also included. - [Read Raf-1 Kinase Assay]

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Run-on transcription monitors the regulation of transcription in isolated nuclei. Protocol includes: Preparation of Nuclei, Run-On Transcription Reaction, Alkaline Hydrolysis of Run-Off Transcripts, Preparation of Filters and Hybridization. Solutions included: DNase Solution, Hybridization Mix, Denhardt Solution (100X) etc. - [Read Run- On Transcription Protocol]

Category: Cell Biology and Cell Culture: Cell Culture Techniques: Cell Line Preservation Freezing

Thawing & Freezing Protocols for Insect Cells (BV - for Baculovirus ) - eBioscience. Freezing Protocol and Thawing Protocols included. - [Read Thawing & Freezing Protocols for Insect Cells (BV - Baculovirus ) - eBioscience]

Category: Molecular Imaging: Confocal Microscopy Protocols and Information

Zeiss Meta Confocal Microscopy Imaging Instructions. Operating Instructions with photos included. Keck Imaging Center. - [Read Zeiss Meta Confocal Microscopy Imaging Instructions]