Chromosomal DNA extraction after restriction endonuclease treatments: a study by microdensitometry and electrophoresis.
The appearance of G- and C-banding patterns on metaphase chromosomes from the Don cell line (Cricetulus griseus) was accompanied by extraction of chromosomal DNA. A microdensitometric technique was employed to measure the loss of DNA produced by Hin dIII, Hae III and Eco RI endonuclease digestion on fixed chromosomes, from 0.5 to 24 h of treatment. Agarose gel electrophoresis was carried out to estimate the size of the DNA fragments extracted after digestion by Alu I and the restriction endonucleases already mentioned. Results obtained show that Alu I and Hae III, which possess 4 base pair recognition sequences, caused higher DNA extraction than enzymes with recognition targets of 6 base pairs (Hin dIII and Eco RI). Incubation buffers induced G-banding patterns which were accompanied by DNA extraction, although this loss was lower than that produced by endonuclease treatments.
González-Tizón
A M
AM
Departamento de Biología Celular y Molecular, Universidad de La Coruña, Spain.
Martínez-Lage
A
A
Méndez
J
J
D000818:Animals; D001399:Azure Stains; D002021:Buffers; D002460:Cell Line; D002875:Chromosomes / Q000737:chemistry* / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000187:drug effects / Q000302:isolation & purification*; D003720:Densitometry; D015246:Deoxyribonuclease EcoRI / Q000494:pharmacology*; D015247:Deoxyribonuclease HindIII / Q000494:pharmacology*; D004587:Electrophoresis, Agar Gel; D007621:Karyotyping; D008677:Metaphase
D001399:Azure Stains; D002021:Buffers; D004247:DNA; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII