How and where to obtain information on Neurospora
How to procure Neurospora strains for research and teaching
How to choose wild type laboratory strains for reference and special uses
How to certify that Neurospora is non-hazardous
How to avoid pitfalls and keep out of trouble in the Neurospora lab
How to avoid contamination by airborne conidia
How to deal with mite infections and to minimize their occurrence
How to make very small inocula
How to prepare culture media
How to measure and monitor linear growth
How to prepare and use gradient plates
How to convert wild-type spreading growth to colonial
Conidiation
How to maximize the production of macroconidia
How to obtain microconidia
How to convert wild-type spreading growth to colonial
How to plate conidia and ascospores
Simply-prepared filters for conidial suspensions
How to do replica plating
How to prepare and use gradient plates
How to make disposable spreaders from long-nose Pasteur pipets
Media
How to choose and prepare media
How to
color-code media
Stock preservation
How to
preserve stocks
How to deposit materials to the FGSC
How to stain and count nuclei
How to use hematoxylin for cytological studies
How to stain meiotic chromosomes using acriflavine
How to prepare aceto-orcein squashes, especially for pachytene chromosomes
How to prepare ascus rosettes for microscopic examination
How to
separate chromosomes and restriction fragments using pulsed field gel
electrophoresis
How to do film and digital imaging of microscopic preparations
How to
obtain naked nuclei extruded from the cell
How to minimize bubble-ascus abortion in crosses for cytology
How to use GFP-tagged proteins in cell biology
How
to work with slime and other cell wall-deficient strains
How to prepare mitochondria from mycelia at different growth phases
How to prepare
vacuolar membranes and other membrane fractions
How
to use cryofixation and freeze substitution for ultrastructural studies of
hyphae
How to analyze and quantify conidial anastomosis tube (CAT)-mediated cell fusion
How to distinguish conidial anastomosis tubes (CATs) from germ tubes
Making crosses
How to choose wild type laboratory strains for reference and special uses
How to determine mating type
How to use fluffy testers for determining mating type and for other applications
How to make a cross
How to make large
numbers of crosses rapidly
How to enhance the formation of perithecia and the production of progeny by
poorly fertile crosses
How to use helper strains for maintaining and crossing handicapped recessive
mutants, for forcing and resolving heterokaryons, and for determining
heterokaryon compatibility
How to obtain progeny as random ascospore progeny
How to obtain asci as unordered groups of ascospores ejected from the
perithecium
How to obtain unbroken linear asci that are
extruded individually from the perithecium
How to activate ascospores and optimize the recovery of progeny
How to make tapered platinum-iridium needles and use them for isolating
ascospores
How to use per-1 to establish parentage in reciprocal crosses
How to obtain uncontaminated progeny from crosses involving per-1
How to use genetic methods for detecting linkage
How to
use alcoy for linkage group assignment
How to determine gene order using 3-point crosses
How to use multiply marked multicent strains for linkage group assignment of
genes and rearrangement breakpoints
How
to use duplication-generating rearrangements in mapping
How to use RFLP for mapping cloned genes
How to use Spore killer for distinguishing 1st- and 2nd-division segregation
with unordered asci
How to use
auxanograms to identify nutritional requirements
How to optimize carotenoid production
How to assay light responses in Neurospora
How
to work with slime and other cell wall-deficient strains
How to use fluffy testers for determining mating type and for other applications
How to distinguish mitochondrial mutations from nuclear mutations
How to distinguish a Spore killer from alternative causes of ascospore abortion
How to test for complementation between mutant strains
How to determine whether a strain will undergo senescence
How to wash agar
How to test heat, osmotic, and oxidative tolerance of Neurospora conidia and
young mycelia
Stock
building. Genetic manipulation
How to inbreed for increased isogenicity
How to use asci for obtaining double mutants of genes that show epistasis of are
phenotypically similar
How to introgress genes from one species to another
How to make a gene deletion (Knock Out)
How to enrich for mutations
How to determine recessive-lethal mutation rates
How to name genes and assign symbols to previously unnamed loci
How to use the mtr gene as a selectable
marker for loss or gain of function
How to
do replica plating
How to measure mitotic instability of segmental duplications
How to use chemical mutagenesis
How
to determine whether a gene is essential for survival
How to use spot tests to determine mutagen sensitivity
How to use
UV for mutagenesis
How to
Prepare Spheroplasts
How to transform by electroporation
How to make heterokaryons in Neurospora crassa
How to test for complementation between mutant strains
How to separate the components of a heterokaryon
How to identify and score genes that confer vegetative (heterokaryon)
incompatibility
How to demonstrate barrage in Neurospora
How to use helper strains for maintaining and crossing handicapped recessive
mutants, for forcing and resolving heterokaryons, and for determining
heterokaryon compatibility
How to recognize and diagnose chromosome rearrangements
How to obtain and recognize partial-diploid strains that are duplicated for
known chromosome segments
How to measure mitotic instability of segmental duplications
How
to use duplication-generating rearrangements in mapping
How to prepare aceto-orcein squashes, especially for pachytene chromosomes
How to distinguish a Spore killer from alternative causes of ascospore abortion
How to sample natural populations
How to determine the species of a wild-collected isolate
How to introgress genes from one species to another
How to distinguish a Spore killer from alternative causes of ascospore abortion
How to use fluffy testers for determining mating type and for other applications
MOLECULAR BIOLOGY
Nucleic acids
How to isolate DNA:
Large scale
How to isolate mRNA
How to carry out a Northern hybridization
How to amplify DNA from macroconidia
Proteins
How to isolate nondenatured protein extracts for western
analysis
How to isolate proteins
How to
immunoblot proteins
How to use immunodepletion to study quantitative relationships of proteins in a
complex
How to do
immunoprecipitation
How to
cross-link proteins
How to analyze light signal transduction in vitro
Organelles
How to prepare
vacuolar membranes and other membrane fractions
How to prepare mitochondria from mycelia at different growth phases
How to
separate chromosomes and restriction fragments using pulsed field gel
electrophoresis
How to
obtain naked nuclei extruded from the cell
Methods from the Fungal Genetics Newsletter
Methods from the Neurospora Newsletter (prior to 1996)
Please contact Dr. D. Bell-Pedersen to participate in the assembly of the complete Neurospora protocol guide.
Last modified 9/9/07 KMC