procedure Protocols

Category: DNA Protocols: DNA Extraction Protocols

DNA extracted from peripheral blood leucocytes using 3mls of whole blood. Modification of a salting out procedure as described by Miller et al., (1988). The DNA Laboratory, Medical School, Malta. - [Read DNA Extraction From Blood Leukocytes]

Category: DNA Protocols: DNA Extraction Protocols

Cheek cells obtained by rinsing the mouth commercial mouth wash solution. Mouth wash is then discarded into a sterile conical tube and sent to the lab. Based on Salting out procedure. DNA Laboratory, Medical School, Malta. - [Read DNA Extraction from Cheek Cells]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

Protocol describing extraction of DNA from Rhizobium. - [Read DNA Extraction Procedure (CsCl) - Rhizobium]

Category: Cytogenetics Protocols: DIG Biotin Labeled Probe Protocols

Protocol describes here a high sensitivity indirect detection procedure for DIG-labeled hybridization probes. The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization. Includes: In situ hybridization with DIG-labeled probes; Detection of DIG-labeled probes; Fluorescence microscopy. - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System]

Category: Nucleic Acid Modification Protocols: DIG Biotin Labeled Probes Protocols

Protocol describes a high sensitivity indirect detection procedure for DIG-labeled hybridization probes.  The procedure uses the components of the HNPP Fluorescent Detection Set to form a fluorescent precipitate of HNPP (2-hydroxy-3-naphthoic acid-2’-phenylanilide phosphate) and Fast Red TR at the site of hybridization.  This procedure can be used to detect single copy sequences as small as 1 kb on human metaphase chromosomes. 
    - [Read DNA In Situ Hybridization with an Alkaline Phosphatase-Based Fluorescent Detection System Protocol]

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols: Genomic DNA Isolation Protocols

In this protocol, the DNA-binding capacity of Wizard MagneSil particles is used to capture and release a consistent amount of DNA (100 ng) across a wide range of samples.  At the end of the procedure, the DNA is eluted into 100 µl Elution Buffer to give a final concentration of 1 ng/µl, relieving the need for postpurification DNA quantitation. - [Read DNA IQ Isolation of Genomic DNA from Stains and Buccal Swabs Protocol]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

The original maize DNA miniprep protocol is used extensively for many plant species and different tissues. This slightly modified version is acceptable for most DNA extractions. The procedure has the advantage of isolating DNA from plant material very rapidly. The procedure requires a table-top drill-press (mechanized homogenizer). - [Read DNA Microprep Isolation from Plants Protocol]

Category: Plant Biology Protocols: Plant DNA RNA Isolation Protocols

The original maize DNA miniprep protocol is used extensively for many plant species and different tissues. This slightly modified version is acceptable for most DNA extractions. The procedure has the advantage of speed and its use of inexpensive reagents. - [Read DNA Miniprep Isolation from Plants Protocol]

Category: DNA Protocols: DNA Sequencing Protocols

Protocol describes a simple procedure for purifying sequencing reactions using the MagneSil particles from Promega. - [Read Dye Terminator Cleanup for Automated DNA Sequencing Reactions Protocol]

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Protocol describes a procedure measuring cyclooxygenase activity by quantifying PGE2 produced by enzymatic conversion of arachidonic acid, in the presence or absence of potential inhibitors.  This high-throughput method has the advantage that it directly measures cyclooxygenase activity and requires little enzyme. - [Read ELISA Method Measure Inhibition COX Enzymes]

Category: ELISA Protocols

The cyclooxygenase (COX) reaction can be monitored by measurement of oxygen consumption, peroxidase co-substrate oxidation or prostaglandin (PG) detection. This protocol describes a procedure measuring cyclooxygenase activity by quantifying PGE2 produced by enzymatic conversion of arachidonic acid, in the presence or absence of potential inhibitors. - [Read ELISA Method to Measure Inhibition of the COX Enzymes Protocol]

Category: ELISA Protocols

Method and ELISA Procedure for Measuring Serum Antibody Titer. Komabiotech - [Read ELISA Procedure for Measuring Serum Antibody Titer]

Category: Radioactivity: Autoradiography

Gel Autoradiography Protocols. Drying the gel, exposure, and autoradiography. Protein Fixation and Staining (Optional). ENLIGHTNING - General Procedure for Gel Autoradiography Enhancement. PerkinElmer - [Read ENLIGHTNING - General Procedure for Gel Autoradiography Enhancement]

Category: General Research Protocols and Methods: Centrifuge Use and Centrifugation

Excellent Centrifuge Safety Use Manual. 30 pages. Training, Use, Maintenance, Records and Responsibility, Mechanical Breakdown, Emergency Procedure, Specific Safety Instructions Beckman Sorvall and Eppendorf. HSE UBC. - [Read Excellent Centrifuge Safety Use Manual PDF]

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: Plant Biology Protocols: Arabidopsis Protein Peptide Protocols

The simplest way to analyze proteins is in unfractionated extracts. However, it is often desirable to fractionate proteins, e.g, by size. This procedure extracts total protein from Arabidopsis samples. Typical yields are ~2-3 mg/ml (using rosette leaves) or 6-8 mg/ml (using young seedlings). - [Read Extraction of Total Protein from Arabidopsis Protocol]

Category: Cytogenetics Protocols: Fluorescent In Situ Hybridization FISH Protocol

Method assesses cellular mRNA transcripts in tissue sections and cell cultures using unique short anti-sense primers directed against sequences in particular protein(s). The unlabeled synthetic cDNA oligonucleotide primers are extended complementary to a sense mRNA transcript using reverse transcriptase and labeled through incorporation of a fluorescent-labeled dUTP nucleotide base. This procedure provides rapid detection of low abundance mRNA messages that can be related to other cellular.... - [Read Fluorescent In Situ Transcription in Cells and Tissues Protocol]

Category: Cell Biology and Cell Culture: Cell Cycle Protocols: G Phase Protocols

This protocol provides a method for the synchronization of a monolayer culture of CHO cells in G1 using isoleucine deprivation. Since CHO cells can also be adapted to grow in suspension culture, this procedure can be used to obtain larger quantities of cells. When isoleucine is replaced, the cells resume growth and begin to enter S phase ~4 hours later. This method arrests almost 100% of the CHO cells in G1, and upon reversal, leads to rapid recovery of cell growth and very high cell viability. - [Read G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol]

Category: Cell Biology and Cell Culture: Flow Cytometry Protocols: Cell Cycle Analysis Protocols

This protocol provides a method for the synchronization of a monolayer culture of CHO cells in G1 using isoleucine deprivation. Since CHO cells can also be adapted to grow in suspension culture, this procedure can be used to obtain larger quantities of cells. When isoleucine is replaced, the cells resume growth and begin to enter S phase ~4 hours later. This method arrests almost 100% of the CHO cells in G1, and upon reversal, leads to rapid recovery of cell growth and very high cell viability. - [Read G1 Synchronization of CHO Cells by Isoleucine Deprivation Protocol]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

A crude lysate gel assay can be performed to roughly quantitate the DNA in lysates. This is often a valuable time saving step to determine if the phage yield is sufficient to warrant continuing the procedure. - [Read Gel Assay to Determine DNA Content of Phage Lysates Protocol]

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

This procedure capitalizes on the asexual/sexual reproductive cycle of yeast cells. Laboratory yeast strains can be maintained indefinitely by asexual reproduction as one of two haploid mating types, MATa or MAT{alpha}. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array]

Category: Staining Protocols: Gram Staining Protocols

Gram-positive and Gram-negative organisms form a complex of crystal violet and iodine within the bacterial cell during the Gram-staining procedure. Gm+ organisms are thought to resist decolorization by alcohol or acetone because cell wall permeability is markedly decreased when it is dehydrated by these solvents. Thus, the dye complex is entrapped within the cell, resist being washed out by the solvents, and Gm+ bacteria remain purple following this differential stain. - [Read Gram Staining Protocol]

Category: DNA Protocols: SNP Protocols

High-resolution SNP mapping by denaturing HPLC. A SNP mapping procedure that relies on resolving polymorphisms by denaturing HPLC without the necessity of determining the nature of the SNPs. They demonstrate the use of denaturing high-performance liquid chromatography to identify mutations in the candidate genes and to fine-map chromosomal breakpoints. - [Read High-resolution SNP mapping by denaturing HPLC]

Category: Fungi Protocols: Neurospora Protocols

Protocol on how to Isolate Tightly-Coupled Mitochondria from Neurospora crassa mycelium. A Fast and Reproducible Procedure - [Read How to Isolate Tightly-Coupled Mitochondria from Neurospora crassa mycelium Protocol]