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Read the Fine Print
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(Complete Item Description)
- Abstract:
Genomic and plasmid-encoded RAS2 genes from Baker's and brewing yeast strains will be analyzed using a shortened Southern blot procedure employing chemiluminescence. Unique parts of this procedure are the gravity-driven "upside down" transfer and a rotisserie-like hybridization oven, which greatly facilitate shortening the transfer time to 45 minutes and the hybridization step to 60 minutes. Chemiluminescence obviates problems associated with radioisotopes without sacrificing sensitivity: the plasmid-borne RAS gene can be visualized within 15 seconds and the single-copy yeast genomic RAS2 gene, within a minute. Topics for discussion related to this laboratory can include the concepts of RFLPs, evolutionary conservation of DNA sequences and the genetic basis of cancer.
- Subject:
- Science and Technology
- Grade Level:
- Post-secondary
- Collection:
-
Association for Biology Laboratory Education (ABLE)
Read the Fine Print
-
(Complete Item Description)
- Abstract:
Students perform Western blots with polyacrylamide gel electrophoresis of detergent extracts of SV40-transformed cell lines grown in monolayer culture. The gels are blotted onto nitrocellulose and treated with a monoclonal antibody against SV40 T antigen or against the tumor suppressor p53. Blots are then treated with a secondary antibody (anti-mouse IgG conjugated to horseradish peroxidase), and exposed to revealing chemicals and x-ray film. I give one application: induction of p53 by ultraviolet light.
- Subject:
- Science and Technology
- Grade Level:
- Post-secondary
- Collection:
-
Association for Biology Laboratory Education (ABLE)
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