scale Protocols

Category: Molecular Biology Protocols: Carbohydrate Protocols

Solvent partition protocol allows the isolation of gangliosides from small samples and from samples where ganglioside concentrations are low, especially relative to the concentration of potentially contaminating proteins and other large molecular weight species. Stephan Ladisch Director, Center for Cancer and Transplant Biology, Children's National Medical Center, Washington, D.C. - [Read A Method for Micro-Scale Isolation and Purification of Gangliosides]

Category: Cell Biology and Cell Culture: Apoptosis Protocols

A simple technique for quantifying apoptosis in 96-well plates. Quantitative and Large scale assay. Ribble et al 2005. - [Read A simple technique for quantifying apoptosis in 96-well plates]

Category: DNA Protocols: Genomic DNA Library Protocols

Protocol for bacteriophage lamda plant genomic library construction. Includes: Preparation of the insert DNA; Pilot Sau3A Digestion; Full-scale partial digestions. - [Read Bacteriophage Lamda Plant Genomic Library Construction Protocol]

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

The ability to synthesize RNA in the lab is critical to many techniques.Radiolabeled and nonisotopically labeled RNA probes, generated in small scale transcription reactions can be used in blot hybridizations and nuclease protection assays. This article includes information on: Requirements For Transcription, RNA Phage Polymerases, Template Options: Plasmids, PCR Products, Oligonuclotides and cDNA, Sense or Antisense, Conventional Or Large Scale Synthesis, Products for In Vitro Transcription. - [Read Basic Information on In Vitro Transcription]

Category: Cell Organelle Protocols: Nucleus Protocols

Protocol for chromatin isolation by small-scale biochemical fractionation. - [Read Chromatin Isolation by Small-Scale Biochemical Fractionation Protocol]

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: DNA Plasmid Preps Protocols

Protocol for CsCl prep of plasmid DNA. This is a standard large scale prep.  For plasmid DNA which gives a yield of 0.5 1.0 mg. - [Read CsCl Prep of Plasmid DNA Protocol]

Category: Cell Biology and Cell Culture

Flow assays offer visualization of cell adhesion under wall shear stress. Visualization of the different events of cell adhesion can be quantified by selective image acquisition and subsequent image processing. Flow assays are suited for adhesive events which occur very rapidly in a time scale shorter than that of most static adhesion assays. Also, events subsequent to the initial events can be studied such as cell stabilization and spreading giving some insight into the kinetics of cell-cell. - [Read Dynamic Flow Assay for Cell Adhesion in a Parallel Plate Flow Chamber]

Category: E Coli Protocols: E coli Protein Protocols

Protocol for the expression of cloned genes in E. coli using IPTG-inducible promoters. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying IPTG-inducible promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using IPTG-inducible Promoters Protocol]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage {lambda} pL promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage lambda pL Promoter Protocol]

Category: E Coli Protocols: E coli Protein Protocols

Protocol for expression of cloned genes in E. coli using the bacteriophage lambda pL promoter. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage lambda pL promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage lambda pL Promoter Protocol]

Category: E Coli Protocols: E coli Protein Protocols

Protocol for the expression of cloned genes in E. coli using the bacteriophage T7 promoter. Protocol describes how (1) to clone cloned sequences encoding open reading frames in plasmids carrying bacteriophage T7 promoters, (2) to optimize expression of target proteins in transformants carrying these recombinants, and (3) to scale-up production of foreign proteins. - [Read Expression of Cloned Genes in E. coli Using the Bacteriophage T7 Promoter Protocol]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Formamide can be used instead of proteinase K to dissociate bacteriophage {lambda} coat proteins from the viral {lambda}. The procedure is rapid and works best with large-scale preparations of bacteriophage {lambda}. - [Read Extraction of Bacteriophage lambda DNA from Large-scale Cultures Using Formamide Protocol]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

DNA is best isolated from bacteriophage lambda particles by digesting the viral coat proteins with a powerful protease such as proteinase K, followed by extraction with phenol:chloroform. This procedure is easily adapted for use with smaller-scale preparations of bacteriophage lambda. - [Read Extraction of Bacteriophage lambda DNA from Large-scale Cultures Using Proteinase K and SDS Protocol]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

Most manipulations with M13, including preparations of viral stocks and isolation of single- and double-stranded DNAs, begin with small-scale liquid cultures that are infected with an M13 plaque, picked from an agar plate. - [Read Growing Bacteriophage M13 in Liquid Culture Protocol]

Category: Genetics and Genomics: Genotyping Protocols: PCR Genotyping

Developed PCR-based single molecule haplotyping methods that enable both surveys for novel inversion variants, and population-scale genotyping of known inversions - [Read Haplotype-Fusion PCR Protocol]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to isolate DNA; Large Scale. - [Read How to isolate DNA; Large Scale]

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation of mRNA-protein complexes. In this protocol, an antibody targeting an RBP of interest is used to immunoprecipitate the RBP and any interacting molecules from a cell lysate. Reverse transcription followed by PCR is then used to identify individual mRNAs isolated with the RBP. This method focuses on examining an association between a specific RBP-mRNA complex, and it is best suited for a small scale screening of known or putative binding partners. - [Read Immunoprecipitation of mRNA-Protein Complexes Protocol]

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

Protocol for in vitro transcription and translation using the coupled reticulocyte lysate system. This protocol is designed to test random samples on a protein gel. Scale up the reactions accordingly. Protocol includes: Procedure, Solutions, BioReagents and Chemicals and protocol hints. - [Read In Vitro Transcription and Translation Using the Coupled Reticulocyte Lysate System]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for the isolation of Arabidopsis nuclei and measurement of gene transcription rates using nuclear run-on assays. Plant materials are ground in hexylene glycol-based buffers and highly enriched nuclear fractions are obtained using Percoll density gradients. Standard and small-scale protocols are presented, along with a tested method for nuclear run-on assays. The entire process may be completed within 3 days. - [Read Isolation of Arabidopsis Nuclei and Measurement of Gene Transcription Rates Protocol]

Category: BACs Bacterial Artificial Chromosome Protocols

The procedure for isolation of BAC DNA is scaled-up to accommodate 500-ml cultures, which, on average, yield 20-25 µg of purified BAC DNA. - [Read Isolation of BAC DNA from Large-scale Cultures Protocol]

Category: BACs Bacterial Artificial Chromosome Protocols

BAC DNAs are prepared from 5-ml cultures of BAC-transformed cells by a modification of the standard alkaline lysis method (Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation). The yield typically varies between 0.1 and 0.4 µg of BAC DNA. - [Read Isolation of BAC DNA from Small-scale Cultures Protocol]

Category: Protein Protocols: Nuclear Extraction Protocols

Homogenize cells or tissue, Pellet nuclei, Lyse nuclei, Reprecipitate nuclear proteins, dialyze nuclear extract. Hattori M et al., DNA Cell Biol. 1990. Homogenization buffer and dialysis buffer recipes. - [Read Large scale nuclear extract preparation]

Category: Antibody Protocols: Antibody Production Protocols

Large scale production of monoclonal antibodies. Includes: Strategies to minimize mAb production costs; A home-made hollow fibre bioreactor system. - [Read Large Scale Production of Monoclonal Antibodies]

Category: Cell Culture Protocols

Information on the methods used to cultivate large amounts of cells for the purpose of obtaining an endogenous or recombinant product. - [Read Large-Scale Cell Culture Protocol]

Category: Yeast Protocols: Yeast Staining Protocols

Protocol describes our method for preparing cells for immunofluorescence, in which all incubations and washes are performed in microtiter dishes. The protocol can also easily be adapted for preparing cells for immunofluorescence in microfuge tubes. - [Read Large-Scale Immunocytology Protocol]