genetic Protocols

Category: PCR Protocols: AFLP PCR

AFLP® technology was developed initially by Keygene in the early 1990’s. AFLP has become a widespread genetic fingerprinting technology. AFLP video and diagrams. Keygene - [Read AFLP - Genetic Fingerprinting]

Category: PCR Protocols: AFLP PCR

Ulrich G. Mueller and L. LaReesa Wolfenbarger. Amplified fragment length polymorphisms (AFLPs) are polymerase chain reaction (PCR)-based markers for the rapid screening of genetic diversity. AFLP. TREE October 1999 - [Read AFLP genotyping and fingerprinting Review PDF]

Category: Cell Biology and Cell Culture: Apoptosis Protocols: Apoptosis Analysis

An Integrative Procedure for Apoptosis Identification and Measurement. Yingyu Cui Lab/Group: National Key Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences. I have uploaded an integrative procedure through which a relatively satisfactory result can be obtained following a single stage of cell culture and transient cell treatment, then detection with different instruments. This shortens experiment time. - [Read An Integrative Procedure For Apoptosis Identification And Measurement]

Category: Reporter Gene Assays: B-Galactosidase Assay Protocols

Method describes how a crude extract is prepared, and the activity is normalized to the amount of protein assayed. This method is particularly suitable for comparing cells that are grown under very different conditions or that have different genetic backgrounds. - [Read Assay of ß-Galactosidase in Yeast: Assay of Crude Extracts]

Category: Yeast Protocols: Yeast Genetics Protocols: B-Galactosidase Assay Protocols

Protocol describes how a crude extract is prepared, and the activity is normalized to the amount of protein assayed. This method is particularly suitable for comparing cells that are grown under very different conditions or that have different genetic backgrounds. - [Read Assay of ß-Galactosidase in Yeast: Assay of Crude Extracts Protocol]

Category: Fungi Protocols: Aspergillus Protocols

Compendium of protocols for using Aspergillus nidulans in genetic, molecular, and cell biological investigations, originally written for members of my research group. It also summarizes our common growth media and nutritional supplements, many of which originally appeared elsewhere but now are difficult to locate. Includes: Growth and storage of Aspergillus nidulans conidia; Nutritional supplements for our common auxotrophies; Double mutants; Mitotic mapping - assigning genes to chromosomes; etc - [Read Fundamentals of Growth, Storage, Genetics and Microscopy of Aspergillus nidulans Protocols]

Category: PCR Protocols

Method describes how to modify the termini of PCR products by introducing restriction sites and other features.  To reduce the chance of contamination with exogenous DNAs, prepare and use a special set of reagents and solutions for PCR only.  Bake all glassware for 6 hours at 150°C and autoclave all plasticware. - [Read Genetic Engineering with PCR Protocol]

Category: PCR Protocols: PCR Optimization

Method describes how to modify the termini of PCR products by introducing restriction sites and other features.  To reduce the chance of contamination with exogenous DNAs, prepare and use a special set of reagents and solutions for PCR only.  Bake all glassware for 6 hours at 150°C and autoclave all plasticware. - [Read Genetic Engineering with PCR Protocol]

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

Protocol for genetic mapping of Arabidopsis centromeres using PCR-based markers. Includes: DNA markers facilitate genome analysis; Centromere mapping using tetrad analysis; Preparation of DNA from frozen plant tissue. - [Read Genetic Mapping of Arabidopsis Centromeres Using PCR-Based Markers Protocol]

Category: C. elegans Protocols: C. elegans Genetics Protocols

A guide to genetic mapping in C. elegans. Includes: Introduction and basics; Two-point mapping; Three-point mapping; Deficiency and duplication mapping; Basics of SNP mapping; Mapping dominant mutations; Mapping suppressor/enhance mutations; Mapping of synthetic mutations. - [Read Guide to Genetic Mapping in C. elegans]

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

Hairpin-bisulfite PCR will be a powerful tool in studying other processes for which genetic or epigenetic information differs on the two complementary strands of DNA. Laird et al., 2003. PNAS - [Read Hairpin-bisulfite PCR: Assessing epigenetic]

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to use genetic methods for detecting linkage. - [Read How to Use Genetic Methods for Detecting Linkage]

Category: Microinjection Protocols: Microinjection of Proteins Protocols

Activation and inactivation of proteins using photoactivation of caged peptides or proteins offer insights into cellular dynamics not achievable using genetic means. The ability to selectively alter the activity of a specific protein at a defined time and location inside a cell allows the correlation of changes in protein activity and cellular behavior. A caged compound, peptide, or protein is prepared by covalently linking it to a photolabile, protecting group. - [Read Introduction of Caged Peptide/Protein into Cells Using Microinjection Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolation of early neural-fold-stage embryos. - [Read Isolating Postimplantation Embryos: Early Neural-Fold-Stage Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause post-implantation defects. This protocol describes a method for isolating early primitive-streak-stage embryos. - [Read Isolating Postimplantation Embryos: Early Primitive-Streak-Stage Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolating early somite-stage embryos (~8.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Early Somite-Stage Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations which cause postimplantation defects. This protocol describes a method for isolating late primitive-streak-stage embryos (~7.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Late Primitive-Streak-Stage Protocol]

Category: Stem Cell Protocols: Stem Cell Isolation Protocols

Isolation of postimplantation-stage embryos allows one to study normal development as well as genetic mutations that cause post-implantation defects. This protocol describes a method for isolating prestreak-stage embryos (~5.5 days post coitum [dpc]). - [Read Isolating Postimplantation Embryos: Prestreak-Stage Protocol]

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: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Molecular and genetic toxicology studies for in vitro cytogenetics with CHO or human lymphocytes. Includes: G401 Chromosomal Aberrations in Chinese Hamster Ovary (CHO) Cells, G401-2, G401-R, G401-R2, G401-R2H, G402 Chromosomal Aberrations in Human Lymphocytes, G402-2, G402-R, G402-R2, G402-R2H, G403 Sister Chromatid Exchanges in Chinese Hamster Ovary (CHO) Cells. - [Read Molecular and Genetic Toxicology Studies for In Vitro Cytogenetics with CHO or human lymphocytes]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Molecular and genetic toxicology studies for in vivo cytogenetics. Includes: G501 Chromosomal Aberrations in Rodent Bone Marrow Cells, G502 Single-Exposure Bone Marrow Cytotoxicity Assay. - [Read Molecular and Genetic Toxicology Studies for In Vivo Cytogenetics]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Molecular and genetic toxicology studies on gene mutation in mammalian cells In vitro. Includes: G201 Mouse Lymphoma L5178Y/tk+/Cell Gene Mutation Assay; G201-R; G202 Mouse Lymphoma L5178Y/tk+/Cell Gene Mutation Screening Assay; G203 Chinese Hamster Ovary or V79 Cell Mutation Assay at the hprt Locus; G203R. - [Read Molecular and Genetic Toxicology Studies on Gene Mutation in Mammalian Cells In Vitro]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Molecular and genetic toxicology studies on microbial mutagenesis. Includes: G101 Salmonella/Microsome Plate Incorporation Assay (Ames Test), G101-R, G102 Salmonella/Microsome Preincubation Assay (Ames Test), G102-R, G103 Salmonella/E.  Coli Reverse Mutation Assay (using four Salmonella and one E. coli strains), G104 Salmonella/E.  Coli Reverse Mutation Assay (using four Salmonella and two E. coli strains), G105, G106. - [Read Molecular and Genetic Toxicology Studies on Microbial Mutagenesis]

Category: Genetics and Genomics: Genotyping Protocols: PCR Genotyping

Using molecular marker technology in studies on plant genetic diversity. DNA-based technologies: PCR-based technologies Amplified fragment length polymorphisms (AFLPs. Includes: AFLP technology, step by step; DNA digestion and ligation; PCRs and detection; Summarising the technology. - [Read PCR-Based Technologies Amplified Fragment Length Polymorphisms (AFLPs)]

Category: Genetics and Genomics: Genotyping Protocols: PCR Genotyping

Plasmodium vivax is the second most prevalent malaria parasite affecting more than 75 million people each year, mostly in South America and Asia. In addition to major morbidity this parasite is associated with relapses and a reduction in birthweight. Practical PCR genotyping protocols based on polymorphic loci present in two P. vivax genetic markers. - [Read Practical PCR Genotyping Protocols for Plasmodium vivax using Pvcs and Pvmsp1]