ووضع العلامات ، والتهجين ، والبروتوكولات الغسيل تحتوي على بعض التعديلات على الوثيقة التي وضعت من قبل الدكتور جي. Quackenbush بينما في معهد للبحوث الجينوم (TIGR) ، الدكتور شون ليفي في الموارد المشتركة ميكروأري فاندربيلت (VMSR) ، والدكتور في حين تسلب ألبا في معهد بويس تومسون (BTI) جامعة كورنيل.

<ol>
	<li>Lorenz, W. W., Sun, F., Liang, C., Kolychev, D., Wang, H., Zhao, X., Cordonnier-Pratt, M. M., Pratt, L. H., Dean, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Water+stress-responsive+genes+in+loblolly+pine+(Pinus+taeda)+roots+identified+by+analyses+of+expressed+sequence+tag+libraries.">Water stress-responsive genes in loblolly pine (Pinus taeda) roots identified by analyses of expressed sequence tag libraries.</a> Tree Physiol. 26, 1-16 (2006).</li><li>Lorenz, W. W., Dean, J. F. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> unpublished data. , .</li><li>Hegde, P., Qi, R., Abernathy, K., Gay, C., Dharap, S., Gaspard, R., Hughes, J. E., Snesrud, E., Lee, N., Quackenbush, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+concise+guide+to+cDNA+microarray+analysis.">A concise guide to cDNA microarray analysis.</a> Biotechniques. 29, 548-562 (2000).</li><li>Alba, R., Fei, Z., Payton, P., Liu, Y., Moore, S. L., Debbie, P., Cohn, J., D'Ascenzo, M., Gordon, J. S., Rose, J. K. C., Martin, G., Tanksley, S. D., Bouzayen, M., Molly, M., Jahn, M. M., Giovannoni, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ESTs,+cDNA+microarrays,+and+gene+expression+profiling:+tools+for+dissecting+plant+physiology+and+development.">ESTs, cDNA microarrays, and gene expression profiling: tools for dissecting plant physiology and development.</a> The Plant Journal. 39, 697-714 (2004).</li><li>Lorenz, W. W., Simões, M., Miguel, C., Dean, J. F. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+Gene+Expression+changes+in+Pinus+species+using+a+Loblolly+pine+cDNA+microarray.">Analysis of Gene Expression changes in Pinus species using a Loblolly pine cDNA microarray.</a> IUFRO-CTIA 2008 Joint Conference. , (2008).</li><li>Simões, M., Lorenz, W. W., Alba, A., Gonçalves, S., Dean, J., Miguel, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+gene+expression+analysis+during+P.+pinaster+embryo+development.">Global gene expression analysis during P. pinaster embryo development.</a> 7th Plant Genomics European Meeting. , (2008).</li></ol>

PtGen2 هو ، والعرف وضعت مؤخرا [كدنا] [ميكروأري التي تم تصميمها ليتم استخدامها في المقام الأول من قبل المجتمع البحوث loblolly الصنوبر. النتائج الأولية ولدت حتى الآن أظهرت الصفيف إلى أن تكون أداة قيمة في تقييم الأحداث التي تحدث في الترانسكربتي استجابة لضغط الجفاف ، وكذلك في رص...

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		<th>Catalogue Number</th>
		<th>Comment</th>
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<td>Pre-hybridization Buffer</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>5X SSC, 0.1% SDS, 1% BSA</td>
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<tr>
<td>Hybridization Buffer</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>30% formamide, 5X SSC, 5X Denhardt&#x2019;s, 1% PolyA RNA (Invitrogen, POLYA.GF), 0.1% SDS</td>
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<tr>
<td>Wash Solution #1</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>1X SSC, 0.2% SDS, preheat to 43&deg;C</td>
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<td>Wash Solution #2</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.1X SSC, 0.2% SDS</td>
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<tr>
<td>Wash Solution #3</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>0.1X SSC</td>
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<td>Isopropyl Alcohol</td>
<td>Reagent</td>
<td>Sigma Aldrich</td>
<td>34959-2.5L</td>
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<td>352070</td>
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<td>Falcon&#x2122;</td>
<td>352196</td>
<td>Use this or a similar cap placed at the bottom of each 50mL conical tube during centrifugation</td>
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<td>Coplin Jar</td>
<td>&nbsp;</td>
<td>Wheaton&#x2122;</td>
<td>900520</td>
<td>&nbsp;</td>
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<td>Staining Dish & Rack </td>
<td>Other</td>
<td>Wheaton&#x2122;</td>
<td>900200</td>
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<td>Invitrogen&#x2122;</td>
<td>L1014-02</td>
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<td>Turbo DNase</td>
<td>Kit</td>
<td>Ambion</td>
<td>AM1907</td>
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<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>Cy-5 Dye</td>
<td>Reagent</td>
<td>GE&#x2122;</td>
<td>PA25001</td>
<td>&nbsp;</td>
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<td>Cy-3 Dye</td>
<td>Reagent</td>
<td>GE&#x2122;</td>
<td>PA23001</td>
<td>&nbsp;</td>
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<td>LifterSlips&trade;</td>
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<td>Erie Scientific Company&#x2122; </td>
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<td>HybChambers</td>
<td>Equipment</td>
<td>GeneMachines</td>
<td>JHYB200003</td>
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processing the loblolly pine ptgen2 cdna microarray

PtGen2 هو ميزة [كدنا] [ميكروأري 26496 تحتوي على تضخيم الصنوبر loblolly التكنولوجيات السليمة بيئيا. وينتج الصفيف في مختبرنا للاستخدام من قبل الباحثين الذين يدرسون التعبير الجيني في الأنواع الصنوبرية الصنوبر وغيرها. وقد وضعت PtGen2 نتيجة لجهودنا اكتشاف الجينات في loblolly الصنوبر ، وتتألف من سلاسل المحددة أساسا من أنسجة الجذر ، ولكن أيضا من إبرة والخلايا الجذعية ، وقد تم اختباره من قبل 1،2 PtGen2 التهجين المختلفة المستهدفة الصنوبرية CY - صبغ المسمى cDNAs ، باستخدام كل من تضخيمها وتضخيم عدم وضع العلامات ، طرق غير مباشرة ، واختبارها أيضا مع عدد من الشروط التهجين والغسيل. هذا الفيديو يركز على معالجة وتصنيع الشرائح قبل وبعد مرحلة ما قبل التهجين ، وكذلك بعد التهجين ، وذلك باستخدام بعض التعديلات على الإجراءات التي وضعت سابقا. 3،4 شملت أيضا ، في شكل النص فقط ، هي البروتوكولات المستخدمة للجيل ، وضع العلامات وتنظيف الهدف من ليالي [كدنا] ، وكذلك معلومات عن البرمجيات المستخدمة لمعالجة البيانات المصب. طبعت PtGen2 مع العازلة طباعة الشخصية التي تحتوي على تركيزات عالية من الملح يمكن أن يكون من الصعب إزالته تماما. تغسل الشرائح الأولى في حل SDS الدافئ قبل قبل التهجين. بعد التهجين المسبق ، تغسل الشرائح بقوة في العديد من التغييرات من المياه لاستكمال إزالة الأملاح المتبقية. ثم يتم تنظيف LifterSlips ™ ووضعها على الشرائح والمسمى [كدنا] تحميل بعناية وعلى ميكروأري عن طريق العمل الشعري الذي ينص على توزيع العينة حتى عبر الشريحة ، ويقلل من فرصة التأسيس الفقاعة. يتم تهجين من الأهداف إلى الصفيف في 48 درجة مئوية في ظروف الرطوبة العالية. بعد التهجين ، يتم إجراء سلسلة من يغسل قياسية بلغت 53 درجة مئوية ودرجة حرارة الغرفة لفترات طويلة. تجهيز PtGen2 الشرائح باستخدام هذا الأسلوب يقلل من الملح وSDS المستمدة من التحف غالبا ما ينظر إليها عند معالجة مجموعة أقل صرامة. التهجين الأهداف المستمدة من مصادر مختلفة عدة RNA الصنوبرية ، أسفرت عن عدد أقل من هذا البروتوكول تجهيز القطع الأثرية ، الخلفية مخفضة ، وتقديم أفضل الاتساق بين المجموعات التجريبية المختلفة من الالواح.

Watch this Scientific Journal Video about Processing the Loblolly Pine PtGen2 cDNA Microarray at JoVE.com

Processing the Loblolly Pine PtGen2 cDNA Microarray

وضعت [كدنا] [ميكروأري PtGen2 للدراسات التعبير الجيني في loblolly الصنوبر ،<em&gt; P. taeda</em&gt; ، والأنواع الأخرى الصنوبرية. هنا ، ونعرض ما قبل وما بعد التهجين المناولة وغسل التقنيات التي يمكن استخدامها مع هذه المجموعة لانتاج أفضل الاتساق والتحف المنخفض ، وانخفاض الخلفيات.

معالجة Loblolly باين PtGen2 ميكروأري [كدنا]

PtGen2 is a 26,496 feature cDNA microarray containing amplified loblolly pine ESTs.  The array is produced in our laboratory for use by researchers ...

processing-the-loblolly-pine-ptgen2-cdna-microarray

Research

JoVE Journal

Biology

7.3K Views.  University of Georgia (UGA). وضعت [كدنا] [ميكروأري PtGen2 للدراسات التعبير الجيني في loblolly الصنوبر ، P. taeda ، والأنواع الأخرى الصنوبرية. هنا ، ونعرض ما قبل وما بعد التهجين المناولة وغسل التقنيات التي يمكن استخدامها مع هذه المجموعة لانتاج أفضل الاتساق والتحف المنخفض ، وانخفاض الخلفيات.

Video: Processing the Loblolly Pine PtGen2 cDNA Microarray

Gibberella zeae Ascospore Production and Collection for Microarray Experiments.

Fusarium graminearum Schwabe (teleomorph Gibberella zeae) is a plant pathogen causing scab disease on wheat and barley that reduces crop yield and grain quality. F. graminearum also causes stalk and ear rots of maize and is a producer of mycotoxins such as the trichothecenes that contaminate grain and are harmful to humans and livestock (Goswami and Kistler, 2004).

The fungus produces two types of spores. Ascospores, the propagules resulting from sexual reproduction, are the main source of primary infection.  These spores are forcibly discharged from mature perithecia and dispersed by wind (Francl et al 1999). Secondary infections are mainly caused by macroconidia which are produced by asexual means on the plant surface. To study the developmental processes of ascospores in this fungus, a procedure for their collection in large quantity under sterile conditions was required. Our protocol was filmed in order to generate the highest level of information for understanding and reproducibility; crucial aspects when full genome gene expression profiles are generated and interpreted. In particular, the variability of ascospore germination and biological activity are dependent on the prior manipulation of the material. The use of video for documenting every step in ascospore production is proposed in order to increase standardization, complying with the increasingly stringent requirements for microarray analysis. The procedure requires only standard laboratory equipment. Steps are shown to prevent contamination and favor time synchronization of ascospores.

To study the developmental processes of ascospores in Gibberella zeae, a procedure for collection under sterile conditions is filmed in order to generate the highest level of information for protocol description. This should facilitate the reproducibility of the experiment, a crucial aspect when full genome expression profile tests are implemented.

بوغ زقي zeae Gibberella الإنتاج والتحصيل للتجارب ميكروأري.

Fusarium graminearum Schwabe (teleomorph Gibberella zeae) is a plant pathogen causing scab disease on wheat and barley that reduces crop yield and grain ...

The Microfluidic Probe: Operation and Use for Localized Surface Processing

Microfluidic devices allow assays to be performed using minute amounts of sample and have recently been used to control the microenvironment of cells. Microfluidics is commonly associated with closed microchannels which limit their use to samples that can be introduced, and cultured in the case of cells, within a confined volume. On the other hand, micropipetting system have been used to locally perfuse cells and surfaces, notably using push-pull setups where one pipette acts as source and the other one as sink, but the confinement of the flow is difficult in three dimensions. Furthermore, pipettes are fragile and difficult to position and hence are used in static configuration only. 
The microfluidic probe (MFP) circumvents the constraints imposed by the construction of closed microfluidic channels and instead of enclosing the sample into the microfluidic system, the microfluidic flow can be directly delivered onto the sample, and scanned across the sample, using the MFP. . The injection and aspiration openings are located within a few tens of micrometers of one another so that a microjet injected into the gap is confined by the hydrodynamic forces of the surrounding liquid and entirely aspirated back into the other opening. The microjet can be flushed across the substrate surface and provides a precise tool for localized deposition/delivery of reagents which can be used over large areas by scanning the probe across the surface. 
In this video we present the microfluidic probe1 (MFP). We explain in detail how to assemble the MFP, mount it atop an inverted microscope, and align it relative to the substrate surface, and finally show how to use it to process a substrate surface immersed in a buffer.

In this video we present the microfluidic probe1 (MFP). We explain in detail how to assemble the MFP, mount it atop an inverted microscope, and align it relative to the substrate surface, and finally show how to use it to process a substrate surface immersed in a buffer solution.

ودقق ميكروفلويديك : تشغيل واستخدام لتصنيع السطحي المترجمة

Microfluidic devices allow assays to be performed using minute amounts of sample and have recently been used to control the microenvironment of cells. ...

RNA Isolation from Embryonic Zebrafish and cDNA Synthesis for Gene Expression Analysis

Many important and complex laboratory procedures require an input of high quality, intact RNA.  A degraded sample or the presence of impurities can lead to disastrous results in downstream experimental applications.  It is therefore, of utmost importance to use solid techniques with numerous safeguards and quality control checks to ensure a superior sample.  Herein, we detail a protocol to isolate total RNA from whole zebrafish embryos using a commercially available chemical denaturant and subsequent cleanup to remove traces of DNA and impurities using a commercial RNA isolation kit. As RNA is relatively unstable and easily prone to cleavage by RNAses, most protocols assay gene expression using a cDNA product that is directly synthesized from an RNA template. We detail a procedure to convert RNA into the more stable cDNA product using a commercially available kit. Throughout these procedures there are numerous quality control checks to ensure that the sample is not degraded or contaminated.  The end product of these protocols is cDNA that is suitable for microarray analysis, RT-PCR or long-term storage.    

The isolation of high quality, intact RNA is an essential step in many laboratory protocols. Here, we demonstrate RNA extraction from whole zebrafish embryos and cDNA synthesis for subsequent application in various experimental procedures including gene expression microarray analysis.

عزل الحمض النووي الريبي من الجنينية التجميعي واسماك الزرد [كدنا] لتحليل الجينات

Many important and complex laboratory procedures require an input of high quality, intact RNA.  A degraded sample or the presence of impurities can lead ...

Global Gene Expression Analysis Using a Zebrafish Oligonucleotide Microarray Platform

Gene microarray technology permits quantitative measurement and gene expression profiling of transcript levels on a genome-wide basis. Gene microarray technology is used in numerous biological disciplines in a variety of applications including global gene expression analysis in relation to developmental stage, to a disease state, and in toxic responses. Herein, we include a demonstration of global gene expression analysis using a comprehensive zebrafish-specific oligonucleotide microarray platform. The zebrafish expression microarray platform contains 385,000 probes, 60 base pairs in length, interrogating 37,157 targets with up to 12 probes per target. For this platform, all cDNA and genomic information available for the zebrafish was collected from various genomic databases including Ensembl (http://www.ensembl.org), VEGA (http://vega.sanger.ac.uk), UCSC (http://genome.ucsc.edu), and ZFIN (http://www.zfin.org). As a result this expression array provides complete coverage of the current zebrafish transcriptome. The zebrafish expression microarray was printed by Roche NimbleGen (Madison, WI). This technical demonstration includes the fluorescent labeling of a cDNA product, hybridization of the labeled cDNA product to the microarray platform, and array scanning for signal acquisition using the one color analysis strategy.  

Gene microarrays are powerful tools in gene expression profiling at a genome-wide level. This technology has application in a variety of biological disciplines including developmental biology and toxicology. In this video, we detail a protocol for global gene expression analysis using a comprehensive oligonucleotide microarray platform for the zebrafish.

تحليل التعبير الجيني العالمي باستخدام ميكروأري قليل النوكليوتيد منهاج اسماك الزرد

Gene microarray technology permits quantitative measurement and gene expression profiling of transcript levels on a genome-wide basis. Gene microarray ...

An Improved Method of RNA Isolation from Loblolly Pine (P. taeda L.) and Other Conifer Species

Tissues isolated from conifer species, particularly those belonging to the Pinaceae family, such as loblolly pine (Pinus taeda L.), contain high concentrations of phenolic compounds and polysaccharides that interfere with RNA purification. Isolation of high-quality RNA from these species requires rigorous tissue collection procedures in the field and the employment of an RNA isolation protocol comprised of multiple organic extraction steps in order to isolate RNA of sufficient quality for microarray and other genomic analyses. The isolation of high-quality RNA from field-collected loblolly pine samples can be challenging, but several modifications to standard tissue and RNA isolation procedures greatly improve results. The extent of general RNA degradation increases if samples are not properly collected and transported from the field, especially during large-scale harvests. Total RNA yields can be increased significantly by pulverizing samples in a liquid nitrogen freezer mill prior to RNA isolation, especially when samples come from woody tissues. This is primarily due to the presence of oxidizing agents, such as phenolic compounds, and polysaccharides that are both present at high levels in extracts from the woody tissues of most conifer species. If not removed, these contaminants can carry over leading to problems, such as RNA degradation, that result in low yields and a poor quality RNA sample. Carryover of phenolic compounds, as well as polysaccharides, can also reduce or even completely eliminate the activity of reverse transcriptase or other polymerases commonly used for cDNA synthesis. In particular, RNA destined to be used as template for double-stranded cDNA synthesis in the generation of cDNA libraries, single-stranded cDNA synthesis for PCR or qPCR's, or for the synthesis of microarray target materials must be of the highest quality if researchers expect to obtain optimal results. RNA isolation techniques commonly employed for many other plant species are often insufficient in their ability to remove these contaminants from conifer samples and thus do not yield total RNA samples suitable for downstream manipulations. In this video we demonstrate methods for field collection of conifer tissues, beginning with the felling of a forty year-old tree, to the harvesting of phloem, secondary xylem, and reaction wood xylem. We also demonstrate an RNA isolation protocol that has consistently yielded high-quality RNA for subsequent enzymatic manipulations.

An Improved Method of RNA Isolation from Loblolly Pine P. taeda L. and Other Conifer Species

Many plant tissues, including phloem and xylem from loblolly pine (Pinus taeda L.), contain high levels of phenolics and polysaccharides that interfere with RNA purification. This presentation discusses techniques for the harvest of field-grown tissues and isolation of RNA of sufficient quality for microarrays and other genomic analyses.

طريقة محسنة لعزل الحمض النووي الريبي من الصنوبر Loblolly ( P. taeda L.) والأنواع الأخرى الصنوبرية

Tissues isolated from conifer species, particularly those belonging to the Pinaceae family, such as loblolly pine (Pinus taeda L.), contain high ...

Microarray Analysis for Saccharomyces cerevisiae

In this protocol, gene expression in yeast (Saccharomyces cerevisiae) is changed after exposure to oxidative stress induced by the addition of hydrogen peroxide (H2O2), an oxidizing agent. In the experiment, yeast is grown for 48 hours in 1/2X YPD broth containing 3X glucose. The culture is split into a control and treated group. The experiment culture is treated with 0.5 mM H2O2 in Hanks Buffered Saline (HBSS) for 1 hour. The control culture is treated with HBSS only. Total RNA is extracted from both cultures and is converted to a biotin-labeled cRNA product through a multistep process. The final synthesis product is taken back to the UVM Microarray Core Facility and hybridized to the Affymetrix yeast GeneChips. The resulting gene expression data are uploaded into bioinformatics data analysis software.

Microarray Analysis for Saccharomyces cerevisiae

In this protocol, gene expression in yeast (Saccharomyces cerevisiae) is changed after exposure to oxidative stress induced by the addition of hydrogen peroxide (H2O2), an oxidizing agent.

تحليل لميكروأري السكيراء الجعوية</em

In this protocol, gene expression in yeast (Saccharomyces cerevisiae) is changed after exposure to oxidative stress induced by the addition of hydrogen ...

Performing Custom MicroRNA Microarray Experiments

microRNAs (miRNAs) are a large family of &tilde; 22 nucleotides (nt) long RNA molecules that are widely expressed in eukaryotes 1. Complex genomes encode at least hundreds of miRNAs, which primarily inhibit the expression of a vast number of target genes post-transcriptionally 2, 3. miRNAs control a broad range of biological processes 1. In addition, altered miRNA expression has been associated with human diseases such as cancers, and miRNAs may serve as biomarkers for diseases and prognosis 4, 5. It is important, therefore, to understand the expression and functions of miRNAs under many different conditions.
Three major approaches have been employed to profile miRNA expression: real-time PCR, microarray, and deep sequencing. The technique of miRNA microarray has the advantage of being high-throughput, generally less expensive, and most of the experimental and analysis steps can be carried out in a molecular biology laboratory at most universities, medical schools and associated hospitals. Here, we describe a method for performing custom miRNA microarray experiments. A miRNA probe set will be printed on glass slides to produce miRNA microarrays. RNA is isolated using a method or reagent that preserves small RNA species, and then labeled with a fluorescence dye. As a control, reference DNA oligonucleotides corresponding to a subset of miRNAs are also labeled with a different fluorescence dye. The reference DNA will serve to demonstrate the quality of the slide and hybridization and will also be used for data normalization. The RNA and DNA are mixed and hybridized to a microarray slide containing probes for most of the miRNAs in the database. After washing, the slide is scanned to obtain images, and intensities of the individual spots quantified. These raw signals will be further processed and analyzed as the expression data of the corresponding miRNAs. Microarray slides can be stripped and regenerated to reduce the cost of microarrays and to enhance the consistency of microarray experiments. The same principles and procedures are applicable to other types of custom microarray experiments.

A simple procedure of performing custom microRNA microarray experiments is described. The steps include isolating RNA, labeling RNA and reference DNA, hybridizing the samples to microarrays, scanning the microarrays, quantifying and analyzing hybridization signals.

تجارب أداء مخصص ميكروأري MicroRNA

microRNAs (miRNAs) are a large family of &tilde; 22 nucleotides (nt) long RNA molecules that are widely expressed in eukaryotes 1.  Complex genomes encode ...

A Sectioning, Coring, and Image Processing Guide for High-Throughput Cortical Bone Sample Procurement and Analysis for Synchrotron Micro-CT

Bone is a dynamic and mechanically active tissue that changes in structure over the human lifespan. The products of the bone remodeling process have been studied substantially using traditional two-dimensional techniques. Recent advancements in X-ray imaging technology via desktop micro-computed tomography (&#181;CT) and synchrotron radiation micro-computed tomography (SR&#181;CT) have allowed for the acquisition of high-resolution three-dimensional (3D) scans of a larger field of view (FOV) than other 3D imaging techniques (e.g., SEM) providing a more complete picture of microscopic structures within human cortical bone. The specimen should be accurately centered within the FOV, however, to limit the appearance of streak artifacts known to impact data analysis. Previous studies have reported procurement of irregularly shaped rectilinear bone blocks that result in imaging artifacts due to uneven edges or image truncation. We have applied a geological sampling protocol (coring) to procure consistently sized cortical bone core specimens for SR&#181;CT experiments from the anterior aspect of human femora. This coring method is efficient and minimally destructive to tissue. It creates uniform cylindrical samples that decrease imaging artifacts by nature of being isometric during rotation and providing a uniform path length for X-ray beams throughout scanning. Image processing of X-ray tomographic data of cored and irregularly shaped samples confirms the potential of the technique to improve visualization and analysis of cortical bone microarchitecture. A goal of this protocol is to deliver a reliable and repeatable method for the extraction of cortical bone cores that is adaptable for various types of high-resolution bone imaging experiments. An overarching goal of the work is to create a standardized cortical bone procurement for SR&#181;CT that is affordable, consistent, and straightforward. This procedure may further be adapted by researchers in related fields who commonly evaluate hard composite materials such as in biological anthropology, geosciences, or material sciences.

We employed a geological (coring) sampling protocol to procure cortical bone specimens of uniform size for SR&#181;CT experiments from the anterior aspect of human femora. This method is minimally destructive, efficient, results in cylindrical specimens that minimize imaging artifacts from irregular sample shapes and improves microarchitectural visualization and analysis.

قسم، Coring، ودليل معالجة الصور لارتفاع الإنتاجية اختبار العظام كورتيكية الشراء وتحليل لSynchrotron الدقيقة CT

Bone is a dynamic and mechanically active tissue that changes in structure over the human lifespan. The products of the bone remodeling process have been ...

توضيح بنية الجليكان وحدوثه باستخدام تنميط بوليمر ميكروأري

Microarray Polymer Profiling (MAPP) for High-Throughput Glycan Analysis

Microarray polymer profiling (MAPP) is a robust and reproducible approach to systematically determine the composition and relative abundance of glycans and glycoconjugates within a variety of biological samples, including plant and algal tissues, food materials, and human, animal, and microbial samples. Microarray technology underpins the efficacy of this method by providing a miniaturized, high-throughput screening platform, allowing thousands of interactions between glycans and highly specific glycan-directed molecular probes to be characterized concomitantly, using only small amounts of analytes. Constituent glycans are chemically and enzymatically fractionated, before being sequentially extracted from the sample and directly immobilized onto nitrocellulose membranes. The glycan composition is determined by the attachment of specific glycan-recognizing molecular probes to the extorted and printed molecules. MAPP is complementary to conventional glycan analysis techniques, such as monosaccharide and linkage analysis and mass spectrometry. However, glycan-recognizing molecular probes provide insight into the structural configurations of glycans, which can aid in elucidating biological interactions and functional roles.

تسليط الضوء على المؤلف: بروتوكول MAPP - تطوير تحليل الجليكان 

Microarray polymer profiling (MAPP) is a high-throughput technique for compositional analysis of glycans in biological samples.

التنميط الدقيق للبوليمر (MAPP) لتحليل الجليكان عالي الإنتاجية

Microarray polymer profiling (MAPP) is a robust and reproducible approach to systematically determine the composition and relative abundance of glycans ...