This method is advantageous for saving the occasional cultures that become contaminated. Yeast contaminated cultures will appear cloudy when slightly shaken and lymphocytes will not cluster together as much as normal. If cultures are suspect, a drop of culture can be streaked on a YPD media plate to check for growth of yeast colonies, or a 5 ml sample can be taken to Barnes Diagnostic Center for identification of yeast strain. - [Read Removal of Yeast Contamination from Lymphoblast Cultures Protocol]
Protocol for RUVKUN antibody staining. Includes: Fixation; To make solution; REDUCING DISULFIDES TO -SH; OXIDIZE -SH GROUPS TO -SO3; TO CHECK THAT WORMS ARE PERMEABLE TO MACROMOLECULES; ANTIBODY INCUBATIONS; PERMANENT SPRINGTIME MOUNTING. - [Read RUVKUN Antibody Staining Protocol]
Protocol for RUVKUN Antibody Staining. Includes: Fixation; To make solution; REDUCING DISULFIDES TO -SH; OXIDIZE -SH GROUPS TO -SO3; TO CHECK THAT WORMS ARE PERMEABLE TO MACROMOLECULES; ANTIBODY INCUBATIONS; PERMANENT SPRINGTIME MOUNTING. - [Read RUVKUN Antibody Staining Protocol]
A Single Stranded Plasmid DNA Isolation Protocol describing the production and isolation of single-stranded DNA (ssDNA) using bacteriophagemid-containing bacteria and helper phage. Infection of the host cells with helper phage allows for packaging of ssDNA into bacteriophage. The ssDNA can then be isolated from phage particles.
DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.
This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.