Direct fluorescence in situ hybridization on human metaphase chromosomes using quantum dot-platinum labeled DNA probes

2015 ◽  
Vol 467 (2) ◽  
pp. 328-333
Author(s):  
Gyoyeon Hwang ◽  
Hansol Lee ◽  
Jiyeon Lee
Keyword(s):  
In Situ Hybridization ◽  
Quantum Dot ◽  
Fluorescence In Situ Hybridization ◽  
Dna Probes ◽  
Metaphase Chromosomes ◽  
Direct Fluorescence

A Simplified Combination of DNA Probe Preparation and Fluorescence in situ Hybridization

1992 ◽  
Vol 47 (9-10) ◽  
pp. 739-747 ◽  
Author(s):  
Dino Celeda ◽  
Ulrich Bettag ◽  
Christoph Cremer
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Dna ◽  
Human Lymphocytes ◽  
Dna Probes ◽  
Dna Probe ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Human Dna

Fluorescence in situ hybridization (FISH) has found widespread applications in cytogenetics. So far the standard protocols for probe amplification (and simultaneous labeling) by PCR, nick translation and in situ hybridization involve different buffer systems leading to a number of time consuming washing steps even before hybridization. In this manuscript we show a fast technique of a close combination of DNA probe preparation and in situ hybridization (ISH). This method was applied to metaphase chromosomes from human lymphocytes fixed on slides. Two specific repetitive DNA probes, the pUC 1.77 DNA probe and the DYZ 1 repetitive DNA fraction were used, amplified and labeled in different ways. Additional experiments with total genomic male human DNA as the DNA probe suggest that this method may be extended to a large variety of other probes. Moreover the ISH technique described does not require toxic denaturing agents, such as formamide.

Fluorescence in situ hybridization (FISH) on maize metaphase chromosomes with quantum dot-labeled DNA conjugates

Chromosoma ◽  
2007 ◽  
Vol 117 (2) ◽  
pp. 181-187 ◽  
Author(s):  
Lu Ma ◽  
Sheng-Mei Wu ◽  
Jing Huang ◽  
Yi Ding ◽  
Dai-Wen Pang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Quantum Dot ◽  
Fluorescence In Situ Hybridization ◽  
Metaphase Chromosomes ◽  
Dna Conjugates

Quantum-Dot-Labeled DNA Probes for Fluorescence In Situ Hybridization (FISH) in the MicroorganismEscherichia coli

ChemPhysChem ◽  
2006 ◽  
Vol 7 (5) ◽  
pp. 1062-1067 ◽  
Author(s):  
Sheng-Mei Wu ◽  
Xiang Zhao ◽  
Zhi-Ling Zhang ◽  
Hai-Yan Xie ◽  
Zhi-Quan Tian ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Quantum Dot ◽  
Fluorescence In Situ Hybridization ◽  
Dna Probes

Karyotypes and Distribution of Tandem Repeat Sequences in Brassica nigra Determined by Fluorescence in situ Hybridization

2017 ◽  
Vol 152 (3) ◽  
pp. 158-165 ◽  
Author(s):  
Gui-xiang Wang ◽  
Qun-yan He ◽  
Jiri Macas ◽  
Petr Novák ◽  
Pavel Neumann ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Fluorescence Intensity ◽  
Tandem Repeat ◽  
Brassica Nigra ◽  
Metaphase Chromosomes ◽  
Repeat Sequences ◽  
Tandem Repeat Sequences ◽  
Black Mustard

Whole-genome shotgun reads were analyzed to determine the repeat sequence composition in the genome of black mustard, Brassica nigra (L.) Koch. The analysis showed that satellite DNA sequences are very abundant in the black mustard genome. The distribution pattern of 7 new tandem repeats (BnSAT13, BnSAT28, BnSAT68, BnSAT76, BnSAT114, BnSAT180, and BnSAT200) on black mustard chromosomes was visualized using fluorescence in situ hybridization (FISH). The FISH signals of BnSAT13 and BnSAT76 provided useful cytogenetic markers; their position and fluorescence intensity allowed for unambiguous identification of all 8 somatic metaphase chromosomes. A karyotype showing the location and fluorescence intensity of these tandem repeat sequences together with the position of rDNAs and centromeric retrotransposons of Brassica (CRB) was constructed. The establishment of the FISH-based karyotype in B. nigra provides valuable information that can be used in detailed analyses of B. nigra accessions and derived allopolyploid Brassica species containing the B genome.

scholarly journals Extending the Useful Life Span of DNA Probes for Fluorescence In Situ Hybridization

BioTechniques ◽  
1999 ◽  
Vol 26 (6) ◽  
pp. 1068-1072
Author(s):  
Allen T. Christian ◽  
Holly E. Garcia ◽  
James D. Tucker
Keyword(s):  
In Situ Hybridization ◽  
Life Span ◽  
Fluorescence In Situ Hybridization ◽  
Dna Probes ◽  
Useful Life

Horse Radish Peroxidase Labelled Oligonucleotides and Tyramide Signal Amplification for Fluorescence in Situ Hybridization to Low Abundant Cytokine Mrnas

1997 ◽  
Vol 3 (S2) ◽  
pp. 203-204
Author(s):  
Mariette van de Corput ◽  
Rob van Gijlswijk ◽  
Mark Bobrow ◽  
Tom Erickson ◽  
Roel Dirks ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Signal Amplification ◽  
Signal To Noise Ratio ◽  
High Specificity ◽  
Tyramide Signal Amplification ◽  
Horse Radish Peroxidase ◽  
Metaphase Chromosomes ◽  
Generation Capacity

In recent years, Tyramide Signal Amplification (TSA) has gained acclaim as a very sensitive detection method for immunocytochemsitry and fluorescence in situ hybridization (FISH). To maximally exploit the great signal generation capacity of TSA in mRNA-FISH, minimizing signals emanating from non-specifically bound nucleic acid probe becomes of prime importance, because a specificity check of the signals observed in the cytoplasm is virtually impossible. We reasoned that utilization of synthetic oligonucleotides (ONTs) in stead of commonly used cDNAs or cRNAs would diminish non-specific probe binding and that direct Horse Radish Peroxidase (HRP) labelling of ONTs and TSA would enable their in situ detection.This approach was first tested in metaphase DNA-FISH using chromosome-specific repeats as targets. Using bifunctional crosslinking chemistry and HPLC, 5’-hexylamino oligonucleotides for chromosome specific simple satellite and alphoid sequences were conjugated to HRP and purified. Following 15 - 20 min of situ hybridization of a single HRP-ONT probe to metaphase chromosomes and a direct flurochrome-tyramide detection step, such repeat targets could be visualized with high specificity and excellent signal-to noise ratio.

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