Shimane Journal of Medical Science

Shimane University Faculty of Medicine
ISSN :0386-5959(in print)
ISSN :2433-2410(online)

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Shimane Journal of Medical Science 13
1992 発行

Investigation on the Non-Radioactive Labeling with Digoxigenin-11-2'-Deoxyuridine-5'-Triphosphate for Southern Blot Analysis

Akane, Atsushi
Matsubara, Kazuo
Shiono, Hiroshi
Nakamura, Hiroaki
Hasegawa, Masanori
Kagawa, Masato
Takahashi, Setsunori
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Description
Southern blot analysis using probes labeled with digoxigenin-11-2'-deoxyuridine-5'-triphosphate (digoxigenin-11-dUTP) instead of ^<32>P-labeled deoxyribonucleotide was investigated. A molecular weight standard marker λ-HindIII should have been labeled with pure digoxigenin-11-dUTP instead of the DIG DNA labeling mixture containing both digoxigenin-11-dUTP and 2'-deoxythymidine-5'-triphosphate. For the labeling of probes, nick translation using pure digoxigenin-11-dUTP was more suitable than random primer extension using the labeling mixture. Furthermore, some modifications on stringency were discussed to identify hybridization signals of the target genes more clearly.
NCID
AA00841586