وأيد هذا العمل من قبل NSF HRD-0630456، وهو CREST NSF البرنامج وNSF هو DMR-0934142.

<ol>
	<li>Sannigrahi, B., Sil, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+and+Characterization+of+&#945;,&#969;-bi[2,4-dinitrophenyl+(DNP)]+poly(2-methoxystyrene)+Functional+Polymers.+Preliminary+Evaluation+of+the+Interaction+of+the+Functional+Polymers+with+RBL+Mast+Cells.">Synthesis and Characterization of &#945;,&#969;-bi[2,4-dinitrophenyl (DNP)] poly(2-methoxystyrene) Functional Polymers. Preliminary Evaluation of the Interaction of the Functional Polymers with RBL Mast Cells.</a> Journal of Macromolecular Science, Part A. 45, 664-671 (2008).</li><li>Gordon, K., Sannigrahi, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+Optically+Active+Helical+Poly(2-methoxystyrene).+Enhancement+of+HeLa+and+Osteoblast+Cell+Growth+on+Optically+Active+Helical+Poly(2-methoxystyrene)+Surfaces.">Synthesis of Optically Active Helical Poly(2-methoxystyrene). Enhancement of HeLa and Osteoblast Cell Growth on Optically Active Helical Poly(2-methoxystyrene) Surfaces.</a> Journal of Biomaterials Science. 2, 2055-2072 (2009).</li><li>Baird, E. J., Holowka, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+Effective+Poly(Ethylene+Glycol)+Architectures+for+Specific+Inhibition+of+Immune+Receptor+Activation.">Highly Effective Poly(Ethylene Glycol) Architectures for Specific Inhibition of Immune Receptor Activation.</a> Biochemistry. 2, 12739-12748 (2003).</li><li>Ramakrisna, S., Fugihara, K., Lim, W. -. E., Ma, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Introductions to Electrospinning and Nanofibers. , (2005).</li><li>Kameoka, J., Craighead, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fabrication+of+Oriented+Polymeric+Nanofibers+on+Planar+Surfaces+by+Electrospinning.">Fabrication of Oriented Polymeric Nanofibers on Planar Surfaces by Electrospinning.</a> Applied Physics Letters. 83, 371-3773 (2003).</li><li>Ramakrishna, S., Lala, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polymer+Nanofibers+for+Biosensor+Applications.">Polymer Nanofibers for Biosensor Applications.</a> Topics in Applied Physics. 109, 377-392 (2007).</li><li>Reuven, D., Sil, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Archetypical+Conductive+Polymer+Structure+for+Specific+Interaction+with+Proteins.">Archetypical Conductive Polymer Structure for Specific Interaction with Proteins.</a> Journal of Macromolecular Science Part A: Pure and Applied Chemistry. , (2012).</li><li>Ogunro, O., Karunwi, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chiral+Asymmetry+of+Helical+Polymer+Nanowire.">Chiral Asymmetry of Helical Polymer Nanowire.</a> The Journal of Physical Chemistry Letters. 1, 704-707 (2010).</li></ol>

الإعلان عن أي تضارب في المصالح.

في هذا التقرير، وقد قدمنا ​​طريقة فعالة لإعداد ألياف النانو biofunctional. زينت ألياف النانو إلى مجموعة وظيفية معينة وهو بروتين لنموذج. للإجراءات ونهج ذكرت في هذه الرسالة هي عامة في طبيعتها، ويمكن استخدامه لإعداد ألياف النانو مزينة أي جماعة وظيفية المطلوب. البلمرة الأني?...

<table border="1" style=";text-align:right;direction:rtl"><tbody><tr><td> اسم كاشف </td><td> شركة </td><td> كتالوج رقم </td></tr><tr><td> الصوديوم المعادن </td><td> سيغما الدريخ </td><td> 282065 </td></tr><tr><td> بينزفينون </td><td> سيغما الدريخ </td><td> 239852 </td></tr><tr><td> 2-methoxystyrene </td><td> سيغما الدريخ </td><td> 563064 </td></tr><tr><td> رباعي هيدرو الفوران </td><td> سيغما الدريخ </td><td> 178810 </td></tr><tr><td> كلوروبنزين </td><td> سيغما الدريخ </td><td> 319996 </td></tr><tr><td> تشارك المركز الوطني الجدران واحد </td><td> سيغما الدريخ </td><td> 704113 </td></tr><tr><td> البوليسترين </td><td> سيغما الدريخ </td><td> 81416 </td></tr><tr><td> السيليكون يفر </td><td> السيليكون كويست الدولي </td><td> 720200 </td></tr><tr><td> زايس FESEM </td><td> شركة كارل زايس </td><td> 60 المتطرف </td></tr><tr><td> Probestation مع بوش ولومب MicroZoom II عالي الأداء مجهر </td><td> باوش والصغرىMB </td><td></td></tr><tr><td> لايكا نظام المسح الضوئي متحد البؤر </td><td> لايكا مايكروسيستمز </td><td> TCS SP2 </td></tr><tr><td> شبه femtoamp Sourcemeter البعيد </td><td> شنيعة الآلات </td><td> 6430 </td></tr><tr><td> Autoranging الرقمية آفومتر </td><td> شنيعة الآلات </td><td> 175A </td></tr><tr><td> حقنة مضخة </td><td> شركة Chemyx </td><td> الانصهار 200 </td></tr><tr><td> زايس المجهر الضوئي </td><td> شركة كارل زايس </td><td> زايس / Axiotech </td></tr></tbody></table>

procedure for fabricating biofunctional nanofibers

Electrospinning وسيلة فعالة لمعالجة ألياف النانو إعداد مزينة المجموعات الوظيفية. يمكن استخدام ألياف النانو مزينة المجموعات الوظيفية لدراسة المواد، العلامات البيولوجية التفاعلات أي بمثابة أجهزة استشعار العوامل البيولوجية المحتملة مع أجهزة الكشف عن جزيء واحد و. وقد وضعنا نهجا فعالا لإعداد البوليمرات الوظيفية حيث وظيفة لديه القدرة على الربط تحديدا مع بروتين النموذج. في نظام نموذجنا، ومجموعة وظيفية هي 2،4-dinitrophenyl (DNP) والبروتين هو مكافحة DNP فريق الخبراء الحكومي الدولي (E المناعي). البوليمر وظيفية، α، ω ثنائية [2،4-dinitrophenyl الكابرويك] [بولي (أكسيد الإثيلين)-B-بولي (2-methoxystyrene)-B-بولي (أكسيد الإثيلين)] (CDNP-PEO-P2MS-PEO- ويتم إعداد CDNP)، عن طريق البلمرة الأنيونية المعيشة. وقد أعد البادئ difunctional المستخدمة في البلمرة عن طريق تفاعل نقل الإلكترون من α-methylstyrene والبوتاسيوم معدن (مرآة). تم إضافة مونومر 2-methoxystyreneأول من البادئ، تليها إضافة لمونومر الثاني، أكسيد الإثيلين، وأخيرا تم إنهاء البوليمر العيش من خلال الميثانول. وα، ω-dihydroxyl البوليمر [HO-PEO-P2MS-PEO-OH] وكان رد فعل مع حمض الكابرويك N-2 ،4-DNP-∈ الأمينية، وذلك بربط DCC، مما أدى إلى تشكيل α، ω ثنائية [ 2،4-dinitrophenylcaproic] [بولي (الإيثيلين أكسيد)-B-بولي (2-methoxystyrene)-B-بولي (أكسيد الإثيلين)] (CDNP-PEO-P2MS-PEO-CDNP). وتميزت البوليمرات التي كتبها FT-IR، 1 H NMR والههلامي (GPC). كانت توزيعات الوزن الجزيئي للبوليمرات الضيقة (1،1-1،2)، وكان يستخدم البوليمرات مع الأوزان الجزيئية أكبر من 50،000 في هذه الدراسة. كانت البوليمرات مساحيق قابلة للذوبان في الصفراء ورباعي هيدرو الفوران. A-للذوبان في الماء CDNP PEO-P2MS-PEO-CDNP / DMEG (جليكول dimethoxyethylene) معقدة تربط ويحقق ثابت ملزم الدولة مع فريق الخبراء الحكومي الدولي حل في غضون ثوان قليلة. الوزن الجزيئي العالي (أي غير قابلة للذوبان المياه حوالي 50،000) CDNتم تجهيز-P-PEO P2MS-PEO-CDNP البوليمرات، التي تحتوي على 1٪ الكربون نانوتيوب جدار واحد (SWCNT) في ألياف النانو electroactive (100 نانومتر إلى 500 نانومتر في القطر) على الركيزة السيليكون. مضان الطيفي يدل على أن فريق الخبراء الحكومي الدولي لمكافحة DNP يتفاعل مع ألياف النانو عن طريق الربط مع المجموعات الوظيفية DNP تزيين الألياف. هذه الملاحظات تشير إلى أن ألياف النانو تبشر functionalized مناسب لتطوير جهاز كشف العلامات البيولوجية.

Watch this Scientific Journal Video about Procedure for Fabricating Biofunctional Nanofibers at JoVE.com

Procedure for Fabricating Biofunctional Nanofibers

يوصف هذا النهج فعالة لإعداد ألياف النانو مزينة المجموعات الوظيفية قادرة على التفاعل مع البروتينات على وجه التحديد. النهج يتطلب أولا إعداد البوليمر functionalized مع مجموعة وظيفية مناسبة. وملفقة البوليمر وظيفية في ألياف النانو التي كتبها electrospinning. يتم دراسة فعالية الربط للألياف النانو مع بروتين متحد البؤر المجهر من قبل.

الإجراء لانتاج ألياف النانو Biofunctional

Electrospinning is an effective processing method for preparing nanofibers decorated with functional groups. Nanofibers decorated with functional groups ...

procedure-for-fabricating-biofunctional-nanofibers

Research

JoVE Journal

Biology

12.5K Views.  Clark Atlanta University. يوصف هذا النهج فعالة لإعداد ألياف النانو مزينة المجموعات الوظيفية قادرة على التفاعل مع البروتينات على وجه التحديد. النهج يتطلب أولا إعداد البوليمر functionalized مع مجموعة وظيفية مناسبة. وملفقة البوليمر وظيفية في ألياف النانو التي كتبها electrospinning. يتم دراسة فعالية الر...

Video: Procedure for Fabricating Biofunctional Nanofibers

A Craniotomy Surgery Procedure for Chronic Brain Imaging

Imaging techniques are becoming increasingly important in the study brain function. Among them, two-photon laser scanning microscopy has emerged as an extremely useful method, because it allows the study of the live intact brain. With appropriate preparations, this technique allows the observation of the same cortical area chronically, from minutes to months. In this video, we show a preparation for chronic in vivo imaging of the brain using two-photon microscopy. This technique was initially pioneered by Dr. Karel Svoboda, who is now a Howard Hughes Medical Institute Investigator at Janelia Farm. Preparations like the one shown here can be used for imaging of neocortical structure (e.g., dendritic and axonal dynamics),  to record neuronal activity using calcium-sensitive dyes, to image cortical blood flow dynamics, or for intrinsic optical imaging studies. Deep imaging of the neocortex is possible with optimal cranial window surgeries. Operating under the most sterile conditions possible to avoid infections, together with using extreme care to do not damage the dura mater during the surgery, will result in successful and long-lasting glass-covered cranial windows.


This video and protocol demonstrate how to implant a glass-covered cranial window in rodents. These preparations can be used for chronic in vivo two-photon imaging of the neocortex over time scales of months. It may also be used for other types of imaging, including optical intrinsic signal imaging.

إجراء جراحة حج القحف لتصوير الدماغ المزمن

Imaging techniques are becoming increasingly important in the study brain function. Among them, two-photon laser scanning microscopy has emerged as an ...

Murine Renal Transplantation Procedure

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell et al over 30 years ago (1) and refined by Zhang et al years later (2), few people in the world have mastered this procedure. In our laboratory we have successfully performed 1200 orthotopic kidney transplantations with &gt; 90% survival rate. The key points for success include stringent control of reperfusion injury, bleeding and thrombosis, both during the procedure and post-transplantation, and use of 10-0 instead of 11-0 suture for anastomoses.
Post-operative care and treatment of the recipient is extremely important to transplant success and evaluation. All renal graft recipients receive antibiotics in the form of an injection of penicillin immediately post-transplant and sulfatrim in the drinking water continually. Overall animal health is evaluated daily and whole blood creatinine analyses are performed routinely with a portable I-STAT machine to assess graft function.

Renal transplantation in mice is a technically challenging procedure that requires careful post-operative care and treatment for success.

فأري إجراء زراعة الكلى

Renal orthotopic transplantation in mice is a technically challenging procedure. Although the first kidney transplants in mice were performed by Russell ...

Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure

Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol for bioluminescent optical imaging of HO-1-luciferase transgenic mice reported in this manuscript provides a rapid in vivo assay of the upregulation of the heme oxygenase-1 (HO-1) gene expression in response to the Gua Sha procedure. HO-1 has long been known to provide cytoprotection against oxidative stress. The upregulation of HO-1, assessed by the bioluminescence output, is thought to represent an antioxidative response to circulating hemoglobin products released by Gua Sha. Gua Sha was administered by repeated strokes of a smooth spoon edge over lubricated skin on the back or other targeted body part of the transgenic mouse until petechiae (splinter hemorrhages) or ecchymosis (bruises) indicative of extravasation of blood from subcutaneous capillaries was observed. After Gua Sha, bioluminescence imaging sessions were carried out daily for several days to follow the dynamics of HO-1 expression in multiple internal organs.

Gua Sha, traditional Chinese therapeutic skin scraping, causes subcutaneous microvascular blood extravasation. We report a protocol of bioluminescence imaging of HO-1-luciferase transgenic mice to demonstrate that Gua Sha upregulates heme oxygenase-1 (HO-1) in multiple organs.

التصوير من تلألؤ بيولوجي Upregulation هيم - 1 أوكسيجيناز في إجراء شا غوا

Gua Sha is a traditional Chinese folk therapy that employs skin scraping to cause subcutaneous microvascular blood extravasation and bruises. The protocol ...

Mammary Epithelial Transplant Procedure

This article describes and compares the fat pad clearance procedure developed by DeOme KB et al.1 and the sparing procedure developed by Brill B et al.2, followed by the mammary epithelial transplant procedure. The mammary transplant procedure is widely used by mammary biologists because it takes advantage of the fact that significant development of the mammary epithelium doesn't occur until after puberty. At 3 weeks of age, growth of the mammary epithelial tree is confined to the vicinity of the nipple and the fat pad is largely devoid of mammary epithelium, but by 7 weeks of age the epithelial ductal tree extends throughout the entire fat pad. Therefore, if this small portion of the fat pad containing epithelium, the region between the nipple and the lymph node, is removed at 3 weeks of age, the endogenous epithelium will never populate the mammary fat pad and the fat pad is described as "cleared". At this time, mammary epithelium from another source can be transplanted in the cleared fat pad where it has the potential to extend mammary ductal trees through out the fat pad. This procedure has been utilized in many experimental models including the examination of tumor phenotype in transgenic mammary epithelial tissue without the confounding effects of genotype on the entire animal3, in the identification of mammary stem cells by transplanting cells in limited dilution4,5, determining if hyperplastic nodules proceed to mammary tumors6, and to assess the effect of prior hormone exposure on the behavior of the mammary epithelium7,8.
Three week old host mice are anesthetized, cleaned and restrained on a surgical stage. A mid-sagittal incision is made through the skin, but not the peritoneum, extending from the pubis to the sternum. Oblique cuts are made through the skin from the mid-sagittal incision across the pelvis toward each leg. The skin is pulled away from the peritoneum to expose the 4th inguinal mammary gland. The fat pad is cleared by removing the fat pad tissue anterior to the lymph node. Epithelium fragments or epithelial cells are transplanted into the remaining cleared fat pad and the mouse is closed.

This article demonstrates the procedure developed by DeOme KB et al. (1959) and the sparing procedure developed by Brill B et al. (2008) for clearing the 4th inguinal mammary fat pad of a pubescent mouse in preparation for transplantation of mammary fragments, mammary epithelial cells, or mammary tumor cells.

زرع الثدي الداخلي طلائي

This article describes and compares the fat pad clearance procedure developed by DeOme KB et al.1 and the sparing procedure developed by Brill B et al.2, ...

A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System

Glucocorticoid stress hormones and their artificial derivatives are widely used drugs to treat inflammation, but long-term treatment with glucocorticoids can lead to severe side effects. Test systems are needed to search for novel compounds influencing glucocorticoid signaling in vivo or to determine unwanted effects of compounds on the glucocorticoid signaling pathway. We have established a transgenic zebrafish assay which allows the measurement of glucocorticoid signaling activity in vivo and in real-time, the GRIZLY assay (Glucocorticoid Responsive In vivo Zebrafish Luciferase activitY). The luciferase-based assay detects effects on glucocorticoid signaling with high sensitivity and specificity, including effects by compounds that require metabolization or affect endogenous glucocorticoid production. We present here a detailed protocol for conducting chemical screens with this assay. We describe data acquisition, normalization, and analysis, placing a focus on quality control and data visualization. The assay provides a simple, time-resolved, and quantitative readout. It can be operated as a stand-alone platform, but is also easily integrated into high-throughput screening workflows. It furthermore allows for many applications beyond chemical screening, such as environmental monitoring of endocrine disruptors or stress research.

We describe the procedure and data analysis of a chemical screening system for glucocorticoid stress hormone signaling using zebrafish larvae: the Glucocorticoid Responsive In vivo Zebrafish Luciferase activitY (GRIZLY) assay. The assay sensitively and specifically detects effects on glucocorticoid signaling by compounds that require metabolization or affect endogenous glucocorticoid production.

Glucocorticoid stress hormones and their artificial derivatives are widely used drugs to treat inflammation, but long-term treatment with glucocorticoids ...

Isolation of Human Hepatocytes by a Two-step Collagenase Perfusion Procedure

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. As such, hepatocytes are an important model for a large variety of research questions. In particular, the use of human hepatocytes is especially important in the fields of pharmacokinetics, toxicology, liver regeneration and translational research. Thus, this method presents a modified version of a two-step collagenase perfusion procedure to isolate hepatocytes as described by Seglen 1.
Previously, hepatocytes have been isolated by mechanical methods. However, enzymatic methods have been shown to be superior as hepatocytes retain their structural integrity and function after isolation. This method presented here adapts the method designed previously for rat livers to human liver pieces and results in a large yield of hepatocytes with a viability of 77&plusmn;10%. The main difference in this procedure is the process of cannulization of the blood vessels. Further, the method described here can also be applied to livers from other species with comparable liver or blood vessel sizes.

A modified two-step collagenase perfusion procedure for isolation of human hepatocytes is described. This method can also be applied to other mammalian livers. The isolated hepatocytes are available in high yield and viability, making them a suitable model for scientific research in areas such as liver regeneration, pharmacokinetics and toxicology.

عزل خلايا الكبد البشرية من قبل خطوتين كولاجيناز الإرواء الداخلي

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. ...

Procedure to Evaluate the Efficiency of Flocculants for the Removal of Dispersed Particles from Plant Extracts

Plants are important to humans not only because they provide commodities such as food, feed and raw materials, but increasingly because they can be used as manufacturing platforms for added-value products such as biopharmaceuticals. In both cases, liquid plant extracts may need to be clarified to remove particulates. Optimal clarification reduces the costs of filtration and centrifugation by increasing capacity and longevity. This can be achieved by introducing charged polymers known as flocculants, which cross-link dispersed particles to facilitate solid-liquid separation. There are no mechanistic flocculation models for complex mixtures such as plant extracts so empirical models are used instead. Here a design-of-experiments procedure is described that allows the rapid screening of different flocculants, optimizing the clarification of plant extracts and significantly reducing turbidity. The resulting predictive models allow the identification of robust process conditions and sets of polymers with complementary properties, e.g. effective flocculation in extracts with specific conductivities. The results presented for tobacco leaf extracts can easily be adapted to other plant species or tissues and will thus facilitate the development of more cost-effective downstream processes for commodities and plant-derived pharmaceuticals.

The design-of-experiments procedure presented here allows the evaluation of different flocculants in terms of their ability to aggregate dispersed particles in plant extracts, thus reducing turbidity and the costs of downstream processing.

إجراءات لتقييم كفاءة المرسب لإزالة فرقت الجسيمات من المستخلصات النباتية

Plants are important to humans not only because they provide commodities such as food, feed and raw materials, but increasingly because they can be used ...

Procedure and Key Optimization Strategies for an Automated Capillary Electrophoretic-based Immunoassay Method

New technologies that utilize capillary-based immunoassays promise faster and more quantitative protein assessment compared to traditional immunoassays. However, similar to other antibody-based protein assays, optimization of capillary-based immunoassay parameters, such as protein concentration, antibody dilution, and exposure time is an important prerequisite to the generation of meaningful and reliable data. Measurements must fall within the linear range of the assay where changes in signal are directly proportional to changes in lysate concentration. The process of choosing appropriate lysate concentrations, antibody dilutions, and exposure times in the human bronchial epithelial cell line, BEAS-2B, is demonstrated here. Assay linearity is shown over a range of whole cell extract protein concentrations with p53 and &#945;-tubulin antibodies. An example of signal burnout is seen at the highest concentrations with long exposure times, and an &#945;-tubulin antibody dilution curve is shown demonstrating saturation. In addition, example experimental results are reported for doxorubicin-treated cells using optimized parameters.

A capillary-based immunoassay using a commercial platform to measure target proteins from total protein preparations is demonstrated. In addition, assay parameters of exposure time, protein concentration, and antibody dilution are optimized for a cell culture model system.

إجراءات واستراتيجيات التحسين الرئيسية لأسلوب الآلي المناعة شعرية المستندة إلى الغرواني الكهربي

New technologies that utilize capillary-based immunoassays promise faster and more quantitative protein assessment compared to traditional immunoassays. ...

A Semiautomated ChIP-Seq Procedure for Large-scale Epigenetic Studies

Chromatin immunoprecipitation followed by sequencing (ChIP-Seq) is a powerful and widely used approach to profile chromatin DNA associated with specific histone modifications, such as H3K27ac, to help identify cis-regulatory DNA elements. The manual process to complete a ChIP-Seq is labor intensive, technically challenging, and often requires large-cell numbers (&#62;100,000 cells). The method described here helps to overcome those challenges. A complete semiautomated, microscaled H3K27ac ChIP-Seq procedure including cell fixation, chromatin shearing, immunoprecipitation, and sequencing library preparation, for batch of 48 samples for cell number inputs less than 100,000 cells is described in detail. The semiautonomous platform reduces technical variability, improves signal-to-noise ratios, and drastically reduces labor. The system can thereby reduce costs by allowing for reduced reaction volumes, limiting the number of expensive reagents such as enzymes, magnetic beads, antibodies, and hands-on time required. These improvements to the ChIP-Seq method suit perfectly for large-scale epigenetic studies of clinical samples with limited cell numbers in a highly reproducible manner.

This paper describes a semiautomated, microscaled ChIP-Seq protocol of DNA regions associated with a specific histone modification (H3K27ac) using a ChIP liquid-handler platform, followed by library preparation using tagmentation. The procedure includes control assessment for quality and quantity purposes and can be adopted to other histone modifications or transcription factors.

إجراء سيميومتوميد ChIP-Seq للدراسات اللاجينية على نطاق واسع

Chromatin immunoprecipitation followed by sequencing (ChIP-Seq) is a powerful and widely used approach to profile chromatin DNA associated with specific ...

تعزيز بقاء ونمو نبات المرموسيت باستخدام نهج التربية اليدوية

Hand-Rearing Method for Infant Marmosets

The common marmoset (Callithrix jacchus) is a small and highly social New World monkey with high reproduction rates, which has been proven to be a compelling non-human primate model for biomedical and neuroscience research. Some females give birth to triplets; however, the parents cannot raise all of them. To save these infants, we have developed a hand-rearing method for raising newborn marmosets. In this protocol, we describe the formula of the food, the time for feeding, the configuration of the temperature and humidity, as well as the adaptation of the hand-reared infants to the colony environment. This hand-rearing method significantly increases the survival rate of marmoset infants (without hand-rearing: 45%; with hand-rearing: 86%) and provides the opportunity to study the development of marmoset infants with similar genetic backgrounds raised in different postnatal environments. As the method is practical and easy to use, we anticipate that it could also be applied to other labs working with common marmosets.

تسليط الضوء على المؤلف: أبحاث Marmoset - النطاق والتحديات 

Here, we describe a hand-rearing method for raising infant marmosets in an animal incubator. This method greatly increases the survival rate of marmoset infants, which provides the opportunity to study the development of marmoset infants with similar genetic backgrounds raised in different postnatal environments.