Texas red 5 dutp

Manufactured by Thermo Fisher Scientific

Sourced in United States, China

Texas Red-5-dUTP is a fluorescent nucleotide analog used in various molecular biology applications. It can be incorporated into DNA or RNA during synthesis or labeling.

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Lab products found in correlation

Alexa fluor 488 5 dutp, by Thermo Fisher Scientific (3 mentions) Texas green 5 dctp, by Thermo Fisher Scientific (2 mentions) Nick translation mix, by Roche (1 mentions) Oligonucleotide probes, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions) Bx53 microscope, by Olympus (1 mentions) Dp80 ccd camera, by Olympus (1 mentions) Digoxigenin 11 dutp, by Roche (1 mentions)

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Texas Red (171 citations)

9 protocols using texas red 5 dutp

Multi-Probe Fluorescent In Situ Hybridization for Octoploid Triticales

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Probes Oligo-1162, Oligo-pSc200 and Oligo-pSc250 were hybridized together to mitotic metaphase chromosomes of octoploid triticales MA and MK in the ND-FISH method. After rinsing the chromosome preparations of MA and MK, GISH was carried out on the same slides with the genomic DNA probe of Kustro. Genomic DNA of rye was labeled with Texas Red-5-dUTP (Invitrogen). Probe labeling and in situ hybridization were conducted with the methods described by Han et al.22 . In addition, sequential ND-FISH was used to analyze wheat-rye 1R disomic addition line 14T-71. Probe Oligo-1162 was hybridized to mitotic metaphase chromosomes of 14T-71, probe Oligo-pTa71-2 was hybridized to the same slide after rising the chromosome preparations. Finally, probes Oligo-pSc119.2-1 and Oligo-pTa535-1 were hybridized to the same slide after the second rising of the chromosome preparations.
Images were taken using an epifluorescence microscope (BX51, Olympus) equipped with a cooled charge-coupled device camera operated with HCIMAGE Live software (version 2.0.1.5) and processed with photoshop CS 3.0.

Fu S., Chen L., Wang Y., Li M., Yang Z., Qiu L., Yan B., Ren Z, & Tang Z. (2015). Oligonucleotide Probes for ND-FISH Analysis to Identify Rye and Wheat Chromosomes. Scientific Reports, 5, 10552.

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Genomic DNA Extraction and Multicolor GISH Analysis

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The total genomic DNA was extracted from fresh leaves using a modified CTAB method (Allen et al., 2006 (link)) with one additional purification step to obtain high-quality DNA. The total genomic DNA of Th. ponticum was labeled with Red-5-dUTP (Invitrogen) by nick translation (Nick Translation Mix, Roche). Genomic DNA of CS was fixed in boiling water for 5 min and used as a blocker at a ratio of 1:300. The GISH procedure was performed as described in Fu et al. (2012) (link). For multicolor GISH (mc-GISH) analysis, Th. bessarabicum DNA was labeled with Alexa Fluor 488-5-dUTP (Invitrogen), and Ps. spicata DNA was labeled with Texas Red-5-dUTP (Invitrogen) (Wang et al., 2019 (link)).
The oligonucleotide probes used to identify wheat and Th. ponticum chromosomes by non-denaturing FISH (ND-FISH) analysis included Oligo-pSc119.2 (6-FAM-5′), Oligo-pTa535 (Tamra-5′) (Tang et al., 2014 (link)), Oligo-44 (6-FAM-5′), and Oligo-D (6-FAM-5′) (Tang et al., 2018 (link)), all of which were synthesized by Shanghai Invitrogen Biotechnology Co. Ltd. (Shanghai, China). Chromosomes were counterstained with 4,6-diamidino-2-phenylindole (DAPI). Fluorescent signals were scanned and photographed with an Olympus BX53 microscope equipped with a Photometrics SenSys CCD DP80 camera (Japan) (Wang et al., 2016 (link)).

Wang Y., Cao Q., Zhang J., Wang S., Chen C., Wang C., Zhang H., Wang Y, & Ji W. (2020). Cytogenetic Analysis and Molecular Marker Development for a New Wheat–Thinopyrum ponticum 1Js (1D) Disomic Substitution Line With Resistance to Stripe Rust and Powdery Mildew. Frontiers in Plant Science, 11, 1282.

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Chromosome Identification in Rye Hybrids

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Sequential fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) were carried out on mitotic chromosome spreads of M8003, Austrian rye, N9116H and N9116M, respectively. Chromosome spreads of materials, probe labeling and in situ hybridization were prepared according to the methods previously described by Han et al. [32 (link)] and Li et al. [33 (link)]. Oligonucleotide probes, Oligo-pTa535 and Oligo-pSc119.2, were 5’ end-labelled with 6-carboxyfluorescein (6-FAM) or 6-carboxytetramethylrhodamine (Tamra), synthesized by Shanghai Invitrogen Biotechnology Co. Ltd. (Shanghai, China), as described by Tang et al. [34 (link)]. The genomic DNA of Austrian rye was labeled with Texas Red-5-dUTP (Invitrogen). Photographs were taken with MetaMorph Research Imaging Software (Molecular Devices, Sunnyvale, Calif.) on Olympus BX61 fluorescence microscope, and then processed with Adobe Photoshop CS 3.0.
For meiotic studies, one or two spike(s) were collected from each three different plants to demonstrate the meiotic process, and anthers with pollen mother cells were fixed in Carnoy’s 6:3:1 (ethanol/acetic-acid/chloroform) fixative, and screened using the conventional acetocarmine procedure according to Li et al. [33 (link)].

Li H., Guo X., Wang C, & Ji W. (2015). Spontaneous and Divergent Hexaploid Triticales Derived from Common Wheat × Rye by Complete Elimination of D-Genome Chromosomes. PLoS ONE, 10(3), e0120421.

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Chromosome Analysis of Rye Using FISH

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FISH and GISH were used to analyze the mitotic metaphase cells of materials used in this study. The genomic DNA of rye ‘Kustro’, Oligo-pSc119.2-1 and Oligo-pTa535-1 [15] (link) were used as probes. The genomic DNA of rye ‘Kustro’ was labeled with Texas Red-5-dUTP (Invitrogen). Oligo-pSc119.2-1 and Oligo-pTa535-1 were 5′ end-labelled with 6-carboxyfluorescein (6-FAM) and 6-carboxytetramethylrhodamine (Tamra), respectively [15] (link). Oligonucleotide probes were synthesized by Shanghai Invitrogen Biotechnology Co. Ltd. (Shanghai, China). The two synthesized probes were diluted by using 1×TE solution and the amount applied was operated according to Tang et al. [15] (link). The chromosome spreads of materials were prepared through the methods described by Han et al. [25] . Probe labeling and in situ hybridization were also operated according to Han et al. [25] . Images were taken using an epifluorescence microscope (BX51, Olympus) equipped with a cooled charge-coupled device camera operated with HCIMAGE Live software (version 2.0.1.5) and processed with photoshop CS 3.0.

Tang Z., Li M., Chen L., Wang Y., Ren Z, & Fu S. (2014). New Types of Wheat Chromosomal Structural Variations in Derivatives of Wheat-Rye Hybrids. PLoS ONE, 9(10), e110282.

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Fluorescent Labeling of Repetitive DNA Sequences

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The cloned repetitive sequences from CS and sequences of pTa71 were labeled with Texas Red-5-dUTP (Invitrogen, Carlsbad, CA, USA). The cloned repetitive sequences from barley were labeled with Alexa Fluor-488-5-dUTP (Invitrogen). Sequences of pTa71 were kindly provided by Professor Fangpu Han, Institute of Genetics and Developmental Biology, Chinese Academy of Science, Beijing, China. Denaturing FISH was carried out according to the procedure described by Tang et al. [8 (link)].

Tang S., Qiu L., Xiao Z., Fu S, & Tang Z. (2016). New Oligonucleotide Probes for ND-FISH Analysis to Identify Barley Chromosomes and to Investigate Polymorphisms of Wheat Chromosomes. Genes, 7(12), 118.

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Fluorescence in situ Hybridization and Multicolor GISH for Chromosome Analysis

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Fluorescence in situ hybridization was conducted using synthetic oligonucleotide probes as described by Tang et al. (2014 (link)) and Zhao et al. (2016 (link)). The probes Oligo-pSc119.2 (green) and Oligo-pTa535 (red) were synthesized and labeled with FAM or TAMRA by Shanghai Invitrogen Biotechnology Co. Ltd (Shanghai, China) and used for FISH as described by Tang et al. (2014 (link)).
For mc-GISH analysis, XM943 DNA was labeled with Texas Red-5-dUTP (Invitrogen) and SY41 DNA was labeled with Texas Green-5-dCTP (Invitrogen) (2014). SY183 DNA was used as the blocker. Chromosomes were counterstained with 4, 6-diamidino-2-phenylindole (DAPI). Fluorescent signals were scanned and photographed with an Olympus BX53 microscope equipped with a Photometrics SenSys CCD DP80 camera (Olympus, Tokyo, Japan). (Wang et al. 2016a (link), 2016b (link)).

Wang Y., Wang S., Jia X., Tian Z., Wang Y., Wang C., Zhang H., Liu X., Zhao J., Deng P, & Ji W. (2021). Chromosome karyotype and stability of new synthetic hexaploid wheat. Molecular Breeding : New Strategies in Plant Improvement, 41(10), 60.

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Multiprobe FISH Karyotyping of AK58

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The method of sequential FISH and Non-denaturing FISH (ND-FISH) with different labeled probes for karyotype analysis of AK58 was mainly performed according to the previously preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this this version posted October 13, 2020. ; https://doi.org/10.1101/2020.09.14.295733 doi: bioRxiv preprint published protocol (Fu et al., 2015) . The probe Sec1 for rye specific secalin was labeled with Texas Red-5-dUTP (Invitrogen) or Alexa Fluor 488-5-dUTP (Invitrogen) using nick translation for FISH (Clarke et al., 1996) . The oligonucleotide probes for ND-FISH with centromeric specific probe CCS1, 18S-45SrDNA probe pTa71, and probe pSc119.2 were referred to Tang et al. (Tang et al., 2014) . The repeats probes pSc200 and 5SrDNA were from the previously published information (Fu et al., 2015; Lang et al., 2019) . The synthetic oligo probes were 5' end-labeled with 6-carboxyfluorescein (FAM) for the green signal and 6-carboxytetramethylrhodamine (Tamra) for the red signal. The slides after FISH and ND-FISH were mounted with Vectashield mounting medium containing 1.5 μg/mL 4, 6diamidino -2-phenylindole (DAPI, Vector Laboratories, Burlingame, CA, USA). The FISH images were captured with an Olympus BX-53 microscope equipped with a DP-80 CCD camera.

Ru Z., Juhász A., Li D., Deng P., Zhao J., Gao L., Wang K., Keeble‐Gagnère G., Yang Z., Li G., Wang D., Bose U., Colgrave M.L., Kong C., Zhao G., Zhang X., Liu X., Cui G., Wang Y., Niu Z., Wu L., Cui D., Jia J., Appels R., & Kong X. (2020). 1RS.1BL molecular resolution provides novel contributions to wheat improvement.

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Multicolor FISH and GISH Analysis

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Fluorescence in situ hybridization (FISH) was completed using synthetic oligonucleotide probes as described by Tang et al. (2014) and Zhao et al. (2016) . Probes Oligo-pSc119.2 (green) and Oligo-pTa535 (red) were synthesized and labelled with FAM or TAMRA by Shanghai Invitrogen Biotechnology Co. Ltd.
were used for FISH as described by Tang et al. (2014) .
For the multicolor-GISH (mc-GISH) analysis, Triticum urartu Thum. DNA was labelled with Texas Red-5-dUTP (Invitrogen) and Ae. tauschii Coss. DNA was labeled with Texas Green-5-dCTP (Invitrogen) (2014). Ae. Speltoides DNA was blocker. Chromosomes were counterstained with 4,6-diamidino-2-phenylindole (DAPI). Fluorescent signals were scanned and photographed with an Olympus BX53 microscope equipped with a Photometrics SenSys CCD DP80 camera (Japan). (Wang et al. 2016) .

Wang Y., Wang S., Jia X., Tian Z., Wang Y., Wang C., Zhang H., Liu X., Zhao J., Deng P., & Ji W. (2021). Chromosome Karyotype and Stability Identification of New Synthetic Hexaploid Wheat.

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Chromosome Spread Preparation and GISH/ND-FISH Analysis

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Seeds were germinated in Petri dishes at 23 °C in the dark until the roots reached a length of 1-2 cm. The roots tips were excised from the germinated seeds and treated with nitrous oxide gas for 2 h. Then, the treated root tips were fixed in ice-cold 90% acetic acid and stored in 70% ethanol at 4 °C until used (Kato et al. 2004) (link). Chromosome spreads of materials were prepared using the methods described by Han et al. (2006) (link). Pollen mother cells (PMCs) were collected from young spikes and fixed in absolute ethanol/ chloroform/glacial acetic acid (6:3:1, v/v/v). Meiotic chromosomes were flattened on a slide in a drop of 1% acetocarmine.
The genomic DNA from Th. intermedium and Ps. spicata was labeled with digoxigenin-11-dUTP (fluorescein isothiocyanate detected by yellow-green fluorescence) (Roche), and the rye DNA was labeled with Texas Red-5-dUTP (Invitrogen). Sheared genomic DNA from CS wheat was used as a blocker. Oligo-pSc119.2-1 and Oligo-pTa535-1 were labeled at the 5 0 end with 6-carboxyfluorescein (green color) and 6-carboxytetramethylrhodamine (red color), respectively. The two oligonucleotide probes were synthesized by Shanghai Invitrogen Biotechnology Co., Ltd. (Shanghai, China). GISH and ND-FISH were performed as described by Han et al. (2006) (link) and Fu et al. (2015) (link), respectively.

Nie L., Yang Y., Zhang J, & Fu T. (2019). Disomic chromosome addition from Thinopyrum intermedium to bread wheat appears to confer stripe rust resistance. Euphytica: Netherlands journal of plant breeding, 215(3).

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