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A COMPARISON OF SUBSTRATES FOR QUANTIFYING THE SIGNAL FROM A NONRADIOLABELED DNA PROBE
被引:17
作者
:
KERKHOF, L
论文数:
0
引用数:
0
h-index:
0
机构:
Scripps Institution of Oceanography, La Jolla
KERKHOF, L
机构
:
[1]
Scripps Institution of Oceanography, La Jolla
来源
:
ANALYTICAL BIOCHEMISTRY
|
1992年
/ 205卷
/ 02期
关键词
:
D O I
:
10.1016/0003-2697(92)90448-G
中图分类号
:
Q5 [生物化学];
学科分类号
:
071010 ;
081704 ;
摘要
:
A method for measuring the amount of a nonradiolabeled DNA probe using four detection substrates is described. In preliminary experiments, digoxygenin-labeled DNA was bound to neutral, nylon membranes and detected with anti-digoxygenin antibodies conjugated to alkaline phosphatase. Four substrates [4-nitrophenyl phosphate, 4-methylumbelliferyl phosphate, AttoPhos, and adamantyl 1, 2-dioxetane phosphate (AMPPD)] were assessed for use in a quantitative hybridization assay. Only AttoPhos and AMPPD were found to have detection limits in the low picogram range and to respond linearly to DNA concentrations ranging from 0 to 1250 pg. In subsequent experiments, a 200-bp DNA probe cloned from the marine bacterium Pseudomonas perfectomarina 23S rRNA gene was hybridized to P. perfectomarina genomic DNA and total RNA. The amount of hybridized probe was determined using AttoPhos. Finally, a digoxygenin-labeled oligonucleotide was probed against genomic DNA. Linearity with respect to DNA concentration was observed using both the 200-bp fragment and the oligonucleotide as probes with a final target detection limit of 166 fg. This study demonstrates the substrate AttoPhos can be used to quantify the amount of nonradiolabeled probe hybridized to target with sufficient sensitivity for very dilute samples, such as environmental samples. © 1992.
引用
收藏
页码:359 / 364
页数:6
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INVESTIGATION OF A QUANTITATIVE POST-HYBRIDIZATION SIGNAL AMPLIFICATION SYSTEM FOR MESSENGER-RNA OLIGODEOXYRIBONUCLEOTIDE INSITU HYBRIDIZATION
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共 29 条
[11]
DEMACARIO EC, 1990, BIOTECHNIQUES, V8, P210
[12]
KINETIC BEHAVIOUR OF CALF-INTESTINAL ALKALINE PHOSPHATASE WITH 4-METHYLUMBELLIFERYL PHOSPHATE
FERNLEY, HN
论文数:
0
引用数:
0
h-index:
0
FERNLEY, HN
WALKER, PG
论文数:
0
引用数:
0
h-index:
0
WALKER, PG
[J].
BIOCHEMICAL JOURNAL,
1965,
97
(01)
: 95
-
&
[13]
PROBES FOR QUANTITATING SUBPICOGRAM AMOUNTS OF HIV-1 RNA BY MOLECULAR HYBRIDIZATION
GILLESPIE, D
论文数:
0
引用数:
0
h-index:
0
GILLESPIE, D
THOMPSON, J
论文数:
0
引用数:
0
h-index:
0
THOMPSON, J
SOLOMON, R
论文数:
0
引用数:
0
h-index:
0
SOLOMON, R
[J].
MOLECULAR AND CELLULAR PROBES,
1989,
3
(01)
: 73
-
86
[14]
NONRADIOACTIVE, PHOTOBIOTIN-LABELED DNA PROBES FOR THE ROUTINE DIAGNOSIS OF BARLEY YELLOW DWARF VIRUS
HABILI, N
论文数:
0
引用数:
0
h-index:
0
HABILI, N
MCINNES, JL
论文数:
0
引用数:
0
h-index:
0
MCINNES, JL
SYMONS, RH
论文数:
0
引用数:
0
h-index:
0
SYMONS, RH
[J].
JOURNAL OF VIROLOGICAL METHODS,
1987,
16
(03)
: 225
-
237
[15]
HANSON RS, 1981, MANUAL METHODS GENER, P328
[16]
INVESTIGATION OF A QUANTITATIVE POST-HYBRIDIZATION SIGNAL AMPLIFICATION SYSTEM FOR MESSENGER-RNA OLIGODEOXYRIBONUCLEOTIDE INSITU HYBRIDIZATION
HOYLAND, JA
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MANCHESTER,SCH MED,DEPT PATHOL SCI,STOPFORD BLDG,OXFORD RD,MANCHESTER M13 9PT,LANCS,ENGLAND
HOYLAND, JA
FREEMONT, AJ
论文数:
0
引用数:
0
h-index:
0
机构:
UNIV MANCHESTER,SCH MED,DEPT PATHOL SCI,STOPFORD BLDG,OXFORD RD,MANCHESTER M13 9PT,LANCS,ENGLAND
FREEMONT, AJ
[J].
JOURNAL OF PATHOLOGY,
1991,
164
(01)
: 51
-
58
[17]
NUCLEIC-ACID HYBRIDIZATION ASSAYS EMPLOYING DA-TAILED CAPTURE PROBES .2. ADVANCED MULTIPLE CAPTURE METHODS
HUNSAKER, WR
论文数:
0
引用数:
0
h-index:
0
机构:
GENE TRAK SYST,FRAMINGHAM,MA 01701
GENE TRAK SYST,FRAMINGHAM,MA 01701
HUNSAKER, WR
BADRI, H
论文数:
0
引用数:
0
h-index:
0
机构:
GENE TRAK SYST,FRAMINGHAM,MA 01701
GENE TRAK SYST,FRAMINGHAM,MA 01701
BADRI, H
LOMBARDO, M
论文数:
0
引用数:
0
h-index:
0
机构:
GENE TRAK SYST,FRAMINGHAM,MA 01701
GENE TRAK SYST,FRAMINGHAM,MA 01701
LOMBARDO, M
COLLINS, ML
论文数:
0
引用数:
0
h-index:
0
机构:
GENE TRAK SYST,FRAMINGHAM,MA 01701
GENE TRAK SYST,FRAMINGHAM,MA 01701
COLLINS, ML
[J].
ANALYTICAL BIOCHEMISTRY,
1989,
181
(02)
: 360
-
370
[18]
KERKHOF L, 1991, THESIS U CALIFORNIA
[19]
DETECTION OF FLAVIVIRUS RNA IN INFECTED-CELLS USING PHOTOBIOTIN-LABELED HYBRIDIZATION PROBES
KHAN, AM
论文数:
0
引用数:
0
h-index:
0
机构:
MONASH UNIV, DEPT MICROBIOL, CLAYTON, VIC 3168, AUSTRALIA
MONASH UNIV, DEPT MICROBIOL, CLAYTON, VIC 3168, AUSTRALIA
KHAN, AM
WRIGHT, PJ
论文数:
0
引用数:
0
h-index:
0
机构:
MONASH UNIV, DEPT MICROBIOL, CLAYTON, VIC 3168, AUSTRALIA
MONASH UNIV, DEPT MICROBIOL, CLAYTON, VIC 3168, AUSTRALIA
WRIGHT, PJ
[J].
JOURNAL OF VIROLOGICAL METHODS,
1987,
15
(02)
: 121
-
130
[20]
RAPID CONFIRMATION OF LISTERIA-MONOCYTOGENES ISOLATED FROM FOODS BY A COLONY BLOT ASSAY USING A DIGOXIGENIN-LABELED SYNTHETIC OLIGONUCLEOTIDE PROBE
KIM, CM
论文数:
0
引用数:
0
h-index:
0
机构:
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
KIM, CM
SWAMINATHAN, B
论文数:
0
引用数:
0
h-index:
0
机构:
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
SWAMINATHAN, B
CASSADAY, PK
论文数:
0
引用数:
0
h-index:
0
机构:
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
CASSADAY, PK
MAYER, LW
论文数:
0
引用数:
0
h-index:
0
机构:
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
MAYER, LW
HOLLOWAY, BP
论文数:
0
引用数:
0
h-index:
0
机构:
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
CTR DIS CONTROL,CTR INFECT DIS,DIV BACTERIAL & MYCOT DIS,MENINGITIS & SPECIAL PATHOGENS BRANCH,ATLANTA,GA 30333
HOLLOWAY, BP
[J].
APPLIED AND ENVIRONMENTAL MICROBIOLOGY,
1991,
57
(06)
: 1609
-
1614
←
1
2
3
→