| Autor: |
Keller GH; Biotech Research Laboratories, Inc., Rockville, Maryland 20850., Cumming CU, Huang DP, Manak MM, Ting R |
| Jazyk: |
angličtina |
| Zdroj: |
Analytical biochemistry [Anal Biochem] 1988 May 01; Vol. 170 (2), pp. 441-50. |
| DOI: |
10.1016/0003-2697(88)90656-2 |
| Abstrakt: |
We have developed a versatile chemical method of attaching hapten moieties onto DNA, for the construction of nonisotopic DNA probes. The DNA is reacted with N-bromosuccinimide at alkaline pH, resulting in bromination of a fraction of the thymine, guanine, and cytosine residues, with adenine modified to a lesser extent. The bromine is subsequently displaced by a primary amino group, attached to a linker arm. The other end of the linker arm has a detectable group preattached to it. We have labeled cloned hepatitis B viral (HBV) DNA with the hapten 2,4-dinitrophenyl (DNP) and used it in combination with a high affinity rabbit anti-DNP antibody, for the detection of hepatitis B DNA by slot blotting. This probe was sensitive enough to specifically detect 1 X 10(-17) mol (1 X 10(6) copies) of HBV DNA in total DNA from human serum. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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