Effect of plating medium and phage storage on mutant frequency and titer in the lambda cII transgenic mutation assay.
Environ Mol Mutagen
; 32(4): 325-30, 1998.
Article
en En
| MEDLINE
| ID: mdl-9882006
We examined several experimental parameters of the lambda cI/cII transgenic mutation assay. In the assay, clear plaque lambda phage mutants are identified in a positive selection scheme following rescue of the lambda/LIZ shuttle vector from frozen tissues of Big Blue" transgenic mice. Mutant frequency and titer of phage from various tissues of control and ENU-treated animals was essentially the same on LB or TB1 plating medium, and storage of isolated DNA at 4 degrees C for up to 4 months did not affect either mutant frequency or titer. Storage of packaged phage for 28 days at 4 degrees C did not affect titer. The mean mutant frequency of packaged phage stored 28 days at 4 degrees C was consistently higher than phage plated the same day as packaging (day 0), though the difference was statistically significant in only two of the four samples tested. Reconstruction experiments in which numerically defined titers of known cII mutants were plated on both G1217 and G1225 E. coli strains and incubated at 37 degrees C or 24 degrees C showed highest titers on G1217 at 37 degrees C. The fraction of the G1217, 37 degrees C titer seen in the other strains and conditions varied widely with the cII mutation.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Bacteriófago lambda
/
Mutación
Límite:
Animals
Idioma:
En
Revista:
Environ Mol Mutagen
Asunto de la revista:
BIOLOGIA MOLECULAR
/
SAUDE AMBIENTAL
Año:
1998
Tipo del documento:
Article
País de afiliación:
Estados Unidos
Pais de publicación:
Estados Unidos