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Isolation and Culture of Human Liver Stem Cells
作者：德尔塔 日期：2022-05-04
Isolation and Culture of Human Liver Stem Cells Materials and Methods 1. Human hepatocytes were isolated from fresh surgical specimens of patients undergoing hepatectomies. Healthy liver tissue (5–20g) was used to isolate hepatocytes by collagenase digestion. 2. Liver tissues were isolated and perfused with 350 ml of a warm (37°C) calcium-free buffer. 3. Liver tissues were digested in Digest Medium at 37°C. This resulted in blanching, softening, and dissociation of hepatic tissue and provided complete digestion of the liver in 10–12 minutes. 4. The hepatocytes were released by mincing and pipetting with a large-bore pipette. 5. The cell suspension was filtered through a sterile 100-μm nylon mesh into a beaker placed on ice, sedimented by centrifugation at 50g for 5 minutes, resuspended, and washed two to three times in cold wash medium. 6. The initial plating consisted of Williams Medium E supplemented with glutamine and with 5% fetal calf serum. 7. Unattached cells were poured off 2–3 hours later and replaced with hepatocyte serum-free medium, a highly modified Chees' medium supplemented with 1.25 μg/cm2 collagen to provide a sandwich matrix. 8. Cultures were re-fed with Hepatozyme-SFM (without collagen) at 24 hours and every 48 hours thereafter. 9. Hepatocytes were seeded at a density of 1.0–1.5 × 105 viable cells, 80% viable cells determined by the trypan blue, per cm2 onto collagen-coated culture plates in Hepatozyme-SFM maintained at 37°C, 5% CO2 for 2 weeks. 10. Human cryopreserved normal hepatocytes obtained were also used. 11. After 2 weeks of culture,
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Identification and expansion of the tumorigenic lung cancer stem cell population
作者：德尔塔 日期：2022-05-04
Identification and expansion of the tumorigenic lung cancer stem cell population Lung cancer contains a rare population of CD133+ cancer stem-like cells able to self-renew and generates an unlimited progeny of non-tumorigenic cells. The current technology allows the establishment of long-term lung cancer stem cell cultures from about one-third of the tumors. 1. Tumor samples were obtained. 2. Surgical specimens were washed several times and left overnight in culture system. DMEM–F12 medium supplemented high doses of penicillin/streptomycin amphotericin B 3. Tissue dissociation was carried out by enzymatic digestion (20 μg/ml collagenase II) for 2 h at 37°C. 4. Recovered cells were cultured at clonal density in serum-free medium. 50 μg/ml insulin 100 μg/ml apo-transferrin 10 μg/ml putrescine 0.03 mM sodium selenite 2 μM progesterone 0.6% glucose 5 mM HEPES 0.1% sodium bicarbonate 0.4% BSA glutamine and antibiotics 20 μg/ml EGF 10 μg/ml bFGF. 5. Flasks non-treated for tissue culture were used to reduce cell adherence and support growth as undifferentiated tumor spheres. 6. The medium was replaced or supplemented with fresh growth factors twice a week until cells started to grow forming floating aggregates. 7. Cultures were expanded by mechanical dissociation of spheres, followed by re-plating of both single cells and residual small aggregates in complete fresh medium. 8. Stem cell medium was replaced with Bronchial Epithelial Cell Growth Medium in tissue culture-treated flasks. 9. Identification: Antibodies used were PE-
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Isolation and Culture of Human Brain Tumor Stem Cell
作者：德尔塔 日期：2022-05-04
Isolation and Culture of Human Brain Tumor Stem Cell The isolation, culture, identification, and purification of stem cells from primary human brain tumors of different phenotypes have marked capacity for proliferation, self-renewal, and differentiation. These cells represents a minority of the tumor cell population and are identified by expression of the cell surface marker the positive CD133. These positive CD133 cells, termed the brain tumor stem cells (BTSCs), which are the expression of neural differentiation markers, and are necessary for the proliferation and self-renewal of the tumor in culture. 1. Tumor samples were obtained from the informed consenting patients. 2. Tumors were washed, acutely dissociated in oxygenated artificial cerebrospinal fluid and subject to enzymatic dissociation. 3. Tumor cells were then resuspended in tumor sphere medium (TSM) • Serum-free neural stem cell medium • Human recombinant EGF (20 ng/ml) • bFGF (20 ng/ml) • Leukemia inhibitory factor (10 ng/ml) • Neuronal Survival Factor (NSF) (10 ng/ml) • N-acetylcysteine (60 μg/ml) 4. Plated at a density of 3 × 106 live cells/60-mm plate. 5. RBCs were removed using lympholyte-M. 6. After primary sphere formation was noted, sphere cells were dissociated and plated in 96-well microwell plates in 0.2 ml volumes of TSM. 7. Final cell dilutions ranged from 200 cells/well to 1 cell/well in 0.2-ml volumes. 8. Cultures were fed 0.025 ml of TSM every 2 days until day 7, when the percentage of wells not containing spheres for each cell plating density was calculated and
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Measurement of primary endothelial cell permeability to fluxes of dextran or albumin
作者：德尔塔 日期：2022-05-04
Measurement of primary endothelial cell permeability to fluxes of dextran or albumin The fluxes of albumin or dextran across vascular endothelial cell (ECs) were evaluated using Transwell chambers. Both molecules were measured for the following reasons: 1) they have different sizes, with albumin having a molecular mass of ∼65 kDa and dextran ∼10 kDa; 2) they may use different transport mechanisms across ECs. In addition to pinocytotic and intercellular transport, albumin transport can be modulated by albumin-binding proteins present on the EC surface. ECs were seeded onto Transwell inserts (6.5-mm diameter and 0.4-µm pore size) coated with 12.5 µg/ml fibronectin. One hour later, 0.05 µCi of 125I-labeled human albumin (Iso-Tex Diagnostics) and 250 µg of FITC-conjugated dextran (10 kDa) were added to the top well, and 100-µl aliquots of samples were taken from the bottom chamber every 15 min for 2 h. The amounts of albumin and dextran in the samples were measured using a gamma counter and a fluorescence plate reader. Each time after taking a sample, 100 µl of medium was added back to the bottom chamber. For each time point, the amounts of albumin and dextran present in the bottom wells as well as the amounts in all the samples removed for measurement were calculated, added up, and plotted. In every experiment, fibronectin-coated filter inserts without ECs were included, and the amounts of dextran and albumin in the bottom wells in each sample were expressed relative to the amounts of these two molecules in
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Primary Culture of Pulmonary Arterial Smooth Muscle Cells
作者：德尔塔 日期：2022-05-04
Primary Culture of Pulmonary Arterial Smooth Muscle Cells 1.Primary cultures of Pulmonary Arterial Smooth Muscle Cells (PASMCs) were isolated from human main pulmonary artery. 2.A segment of the main pulmonary artery was excised and placed in a sterile 10-cm dish containing primary cell culture media. 3.The segment was stripped of adventitia with a sterile forceps. 4.The main pulmonary artery segment was then cut longitudinally to open the vessel, and the endothelial layer was removed by gentle rubbing with a cell scraper. 5.The vessel was then cut into 2-mm segments, inverted, and placed on a collagen-coated 35-mm tissue culture dish. 6.A drop of primary cell culture media containing 10% fetal bovine serum, primary cell culture supplements, antibiotics, and antimycotics was then added, and the cells were grown overnight at 37°C in a humidified atmosphere with 5% CO2-95% air. 7.The next day, an additional 2 ml of complete medium were added. 8.The growth medium was subsequently changed every 2 days. 9.When SMC islands could be observed under the microscope, the tissue segment was removed and the individual cell islands were subcloned. 10.Identity was confirmed as PASMCs by immunostaining (>99% positive) with SMC actin. This was taken as evidence that cultures were not contaminated with fibroblasts or endothelial cells. 11.All cultures for subsequent experiments were maintained in primary cell culture media containing 10% fetal bovine serum, primary cell culture supplements, antibiotics, and antimycotics at 37°C in a humidified atmosphere with
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《Science》发表非损伤微测技术研究Ca2+流速的成果
作者：德尔塔 日期：2022-05-04
D型丝氨酸调节谷氨酸受体基因构成的Ca2+通道 2011年3月17日，葡萄牙里斯本大学José Feijó教授的研究成果在世界知名杂志《Science》以“Research Article”的形式在线发表，中国农业大学资源环境学院的刘来华教授参与了本项研究。细胞内游离Ca2+的增加构成了真核细胞基本的信号转导机制，但是Ca2+通道蛋白如何启动信号一直存在争论。 在这项研究中科学家使用非损伤微测技术（vibrating microelectrodes or NMT）测定了氨基酸刺激后花粉管的Ca2+流动，直接证实了植物中具有和动物中相似的神经传递系统。发现谷氨酸受体类似基因（GLRs）减少了通过质膜的Ca2+内流，进而调节花粉管顶端胞质中的Ca2+浓度梯度，最终影响花粉管的生长和形态建成。此外，敲除花粉管丝氨酸消旋酶（SR1的突变体）后GLRs活性下降，导致生长发生缺陷。这项研究揭示了氨基酸调节雄性配子体和雌蕊组织之间全新的信号转导机制，类似于动物神经系统的常见机制。这不仅是植物学的重大发现，对动物学、医学等其他领域的研究也具有很大的参考价值。 非损伤微测技术（NMT，）是一种活体研究技术，无需标记就可以实时地获取离子和小分子的流速，是研究活体信号转导和神经传递系统的最佳手段之一。José Feijó实验室近年来使用非损伤微测技术在高水平杂志发表了多篇论文。本次发表的论文是该实验室研究工作的代表，也标志着非损伤微测技术的应用越来越广泛，产生的成果越来越有影响力。 关键词：谷氨酸受体类似基因（Glutamate Receptor–Like Genes，GLRs）；钙离子通道（Ca2+ Channel）；花粉管（Pollen Tube）；D型丝氨酸（D-Ser） 参考文献：Michard E et al. Science, DOI: 10.1126/science.1201101 链接：
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超声波传感器对液体进行智能检测
作者：德尔塔 日期：2022-05-04
在实验室自动化中要用到大量的微生物培养用孔板和试管对液体进行分析。采用新的超声传感器，能够快速地、以精确到0.1 mm的精度完成液位高度检测。 实验室也开始越来越多地利用自动化设备来完成试验、检验的任务了，例如：检验分析试剂、试样的吸移等等。而在这些工作中所使用的传感器应该能够精准地进行工作，并提供具有可追溯性的结果，同时还必须具有很高的检测速度。举个例子来说，在一个液体介质检查工位上，很重要的一个步骤就是向很小的玻璃试管灌装检验分析用的液体介质，例如灌入需要分析的血液。在把少量的检测试样灌入到微小的试管中之后，在检验分析开始之前，需要对微小试管中液体介质的液位高度进行检测。这些微小的微生物培养器皿的开口很小，有些微生物培育瓶的口径只有3 mm，但仍然要求传感器能够在最短的时间内精确地检测出瓶内液体介质的容量。当前使用的电容式检测技术还达不到理想的检测精度。一个个独立设置的光电式传感器虽然满足了检测精度的要求，但是价格不菲。 超声传感器是一种完全不与被接触介质接触的检测传感器。这一特性在实验室应用中非常有意义：既保证了传感器不被腐蚀，也保证了被测介质不会出现交叉污染。另外，超声波传感器还对周围环境的湿度和灰尘非常不敏感，从而也保证了它们能够长期、稳定地准确工作。 与光电式传感器相比较，超声式传感器不产生光线，而是利用声波进行检测的，因此它能够可靠地检测不同性质的介质，而且与被测介质的透明度和颜色无关。不论被测介质的粘度高低如何，都不会对检测结果产生影响。 连续工作的超声波传感器向被测对象发射出锥形的声波。这束锥形声波恰好可以进入口径不足10 mm的瓶口，对瓶内的液位高低进行检测。为了避免在使用时受到其他因素的限制，生产厂家与用户密切结合，研发出了新型09系列的超声波传感器。这种新型超声波传感器配备了特殊设计的超声波发射嘴，使发射出去的超声波波束直径更小，能够进入口径3 mm的培育瓶
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接触角测量技术原理及其应用领域
作者：德尔塔 日期：2022-05-04
What is contact angle? Contact angle ,θ , is a quantitative measure of the wetting of a solid by a liquid. It is defined geometrically as the angle formed by a liquid at the three phase boundary where a liquid, gas and solid intersect as shown below: It can be seen from this figure that a low values of contact angle (θ) indicates that the liquid spreads, or wets well, while a high contact angle indicates poor wetting. If the angle θ is less than 90 degrees the liquid is said to wet the solid. If it is greater than 90 degrees it is said to be non-wetting. A zero contact angle represents complete wetting. The measurement of a single static contact angle to characterize the interaction is no longer thought to be adequate. For any given solid/ liquid interaction there exists a range of contact angles which may be found. The value of static contact angles are found to depend on the recent history of the interaction. When the drop has recently expanded the angle is said to represent the ‘advanced’ contact angle. When the drop has recently contracted the angle is said to represent the ‘receded’ contact angle. These angles fall within a range with advanced angles approaching a maximum value and receded angles approaching a minimum value. If the three phase (liquid/solid/vapor) boundary is in actual motion the angles produced are called Dynamic Contact Angles and are referred to as ‘advancing’ and ‘receding’ angles. The difference between ‘advanced’ and ‘advancing’, ‘receded’ and ‘receding’ is that in the static case motion is incipient in the dynamic case motion is actual. D
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Isolation of Human Pulmonary Epithelial Cells
作者：德尔塔 日期：2022-05-04
Isolation of Human Pulmonary Epithelial Cells 1. Pulmonary epithelial cells from human lung tissue were isolated. 2. Lungs were perfused with 10 ml of primary cell medium (supplemented with 1% antibiotics and 2 mM L-glutamine) through the pulmonary artery until they were cleared of blood. 3. Bronchoalveolar lavages were performed using 5 × 1 ml of PBS, 1 mM EDTA to remove alveolar leukocytes. 4. One ml of primary cell isolation solution was instilled into the lungs through the trachea. 5. Lungs were then removed and placed in a sterile tube containing 2 ml of primary cell medium (supplemented with 1% antibiotics and 2 mM L-glutamine) and incubated at 4 °C overnight for enzymatic digestion to occur. 6. Lung tissues were further teased apart in primary cell medium, 10% FCS in a dish. 7. The cell isolate was passed through a 100-μm filter, and pulmonary epithelial cells were purified as follows. 8. Collected cells were counted and incubated for 15 min at 4 °C at the appropriate ratios. 9. Cells were then washed and diluted in primary cell medium, 0.5% FCS. 10. Pulmonary epithelial cells thus obtained were plated on 6-well plate (4 × 105 cells/well) in primary cell medium, 10% FCS.
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环境气体浓度的表示方法及有限空间内的中毒危害
作者：德尔塔 日期：2022-05-04
环境气体浓度的表示有很多种不同的方法，在这里做下简单介绍： WA:时间加权浓度,是污染物在指定时间段内的平均浓度； REL：建议浓度限值，每周四十小时工作制，每天最高十小时TWA允许浓度； PWL：允许的浓度值限制，TLV是阈限值，是TWA的允许浓度，对于每周四十小时，每天八小时的正常工作制来说，工人可以在该浓度下连续工作与气体发生接触而不会产生影响； STEL：短期暴露极限值，定义为十五分钟TWA暴露，在一个工作日的任何时候均不得超过该限值，即使八小时TWA处于限值范围内； IDLH：立即危险浓度，不得超过的最高环境浓度，超过后必须采取高可靠性防毒面具。 有毒有害物质可以是原来就存在于有限空间内的，也可以是作业过程中逐渐积聚的，比较常见的有： (1)苯、甲苯、二甲苯。如在有限空间内进行防腐涂层作业时，由于涂料中含有的苯、甲苯、二甲苯等有机溶剂的挥发，造成有毒物质的浓度逐步增高等。 (2)硫化氢。如清理、疏通下水道、粪便池、窑井、污水池、地窖等作业容易产生硫化氢。 (3)一氧化碳。如在市政建设、道路施工时，损坏煤气管道，煤气渗透到有限空间内或附近民居内，造成一氧化碳积聚，以及在设备检修时，设备内残留的一氧化碳泄漏等。这些有毒有害气体检测要选择有毒气体检测仪。
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L-阿拉伯糖抑制大鼠高脂饮食所致体内脂肪的增加
作者：德尔塔 日期：2022-05-04
L-阿拉伯糖抑制大鼠高脂饮食所致体内脂肪的增加 杨庆，谭壮生，仝国辉，张懿，郑珊 (北京市疾病预防控制中心卫生毒理所，北京100013) 关键词:L-阿拉伯糖;高脂饮食;体内脂肪增加 L-阿拉伯糖作为一种健康的食品添加剂，已大量用于食品加工业中，现已被美国食品药品监督管理局(FDA)和日本厚生省批准列入健康食品添加剂，美国医疗协会也将L-阿拉伯糖列入抗肥胖剂的营养补充剂或非处方药。研究表明，L-阿拉伯糖能够非竞争性地抑制肠内蔗糖酶的活力，调节摄取蔗糖后的血糖效应，从而抑制过多的蔗糖转化体内脂肪，用于防治肥胖 和高血脂等疾病［1-2］。同时有报道表明L-阿拉伯糖可以降低由高糖高脂所引起肥胖兔子脂肪指数［3］。但L-阿拉伯糖对大鼠单纯喂饲高脂饲料引起的体内脂肪堆积的影响鲜见报道。本研究探讨了L-阿拉伯糖对大鼠因喂饲高脂饲料引起的体内脂肪堆积的抑制作用。 1材料与方法 1.1材料L-阿拉伯糖(纯度98%，HPLC);高脂营养饲料:基础饲料80%，猪油10%，蛋黄粉10%。饲料来源:军事医学科学院实验动物中心;饲料合格证号:SCXK-(军)2007-005。 1.2动物清洁级雄性Wistar种大鼠50只，体重140～172 g，由军事医学科学院实验动物中心提供，动物合格证号:SCXK-(军)2007-004。饲养环境:室温20.0～22.5℃，相对湿度为45.5%～50.0%，单笼饲养。动物实验室合格证号:SYXK(京)2008-0001号。 1.3方法选用健康成年雄性大鼠50只，根据体重水平随机分为5个组，即阴性对照组、肥胖模型组及3个受试物剂量组，按人体每日推荐摄入量6 g/人/日计算，3个剂量组剂量分别为低剂量组(500 mg/kg)、中剂量组(1 000 mg/kg)、高剂量组(3 000 mg/kg)，每组10只动物。除阴性对照组给普通饲料外，其他各组均给予高脂饲料喂养。灌胃给予不同剂量的受试物，两 对照组灌胃给予同体积的去离子水，均按1 ml/100 g灌胃给予32 d，灌胃1次/d。实验结束前禁食16 h，称体重后断头处死并解剖称取腹部睾丸及肾周围脂肪。 1.4观察指标每周称体重一次并记录大
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铝依赖的拟南芥离子转运具有低pH和铝响应的特异性
作者：德尔塔 日期：2022-05-04
铝依赖的拟南芥离子转运具有低pH和铝响应的特异性 Aluminum-dependent dynamics of ion transport in Arabidopsis: specificity of low pH and aluminum responses 土壤的酸性是限制植物分布的重要因素，世界上超过40%的耕地是酸性土壤。在酸性土壤中，作物生长受到不同的毒性（H+, Al3+, Mn2+）和营养物质的影响，在这些复杂的因素中，Al3+ 和H+的毒性与植物的生长具有高度的相关性。植物的铝毒性主要是当土壤中的pH低于4.5时的Al3+的作用。因此，为了繁育出更加忍耐酸性土壤的植物，有必要研究H+和Al3+的毒性机制。 澳大利亚的科学家使用非损伤微测技术研究了拟南芥对Al3+和低pH响应的离子转运机制。Al3+毒性早期的症状是抑制了根的生长，随后发现低pH减小了H+内流进入根组织，引起了细胞质的酸化。相反，低pH和Al3+共同胁迫减小了伸长区的H+内流，诱导了成熟区的H+外流，导致了细胞所有区域的碱化。低pH诱导了伸长区根际pH的增加，但是低pH和Al3+共同胁迫导致了所有根部区域的pH下降。低pH胁迫减轻了K+的外流，诱导了质膜的去极化，低pH和Al3+共同胁迫显著减少了质膜去极化。处理60min后，低pH引起了质膜去极化，但是低pH和Al3+共同胁迫引起了质膜的超极化。 这项研究发现了低pH和Al3+毒害对根组织和根际的影响有差异，低pH和Al3+减少了野生型拟南芥根部增加酸性环境中根际pH的能力。这可能为研究植物适应非生物胁迫的不同机制提供了基础。 关键词：非损伤微测技术，低pH值（Low-pH），Al3+，H+，K+ 参考文献：Bose J, et al. Physiologia Plantarum, 2010, 139: 401-412. 全文下载：
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Isolation of human primary airway smooth muscle
作者：德尔塔 日期：2022-05-04
Isolation of human primary airway smooth muscle 1. Primary cultures of human primary airway smooth muscle (ASM) cells were prepared from explants of airway smooth muscle. 2. The human trachea tissue was transported to the laboratory in primary cell culture medium containing 10% fetal calf serum (FCS), primary cell culture supplements, penicillin G (100 u ml−1), streptomycin (100 μg ml−1), amphotericin B (2.5 μg ml−1) and L-glutamine (4 mm). 3. The human trachea tissue was then washed 5 times in the same medium. 4. The trachea muscle was then dissected free of epithelium and connective tissue under sterile conditions. 5. Small (2 × 2 mm) explants of airway smooth muscle were then excised and about 10 explants placed in small dishes. 6. After explants were allowed to adhere, primary cell culture medium, containing FCS, antibiotics, primary cell culture supplements, amphotericin B and L-glutamine, was added to cover the explants. 7. The explants were incubated in humidified 5% CO2/95% air at 37°C. 8. The medium was changed every 3 days. 9. Smooth muscle cells were usually seen about 7 days later. 10. When cells were about to become confluent in some parts of the dish, the explants were removed. 11. Once confluent, cells were trypsinized with 0.25% trypsin and 0.02% EDTA in phosphate-buffered saline (PBS), centrifuged and resuspended in the above medium, counted and plated out in several 75 cm2 flasks and grown to confluence. 12. Cells were then detached with trypsin-EDTA, resuspended in 90% FCS + 10% dimethyl sulphoxide at a density of 106 cells ml−1, frozen in liqui
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ATP生物发光技术的发展与应用
作者：德尔塔 日期：2022-05-04
ATP生物发光技术的发展与应用 ATP是化学物质三磷酸腺苷的简称，存在于所有的生物体中（从微生物到高等动物），ATP在细胞体内主要作用是提供能量。鉴于ATP存在于所有生物体中，利用ATP发光检测仪检测ATP，可以间接地证明生物体的存在。随着食品行业对食品卫生质量要求越来越高，而且ATP生物发光法在检测食品微生物时简单、快速且灵敏度高，因此近年来受到广泛关注。 1 ATP生物发光技术的发展过程 ATP生物发光技术产生于20世纪70年代中期。1983年，Moyer[1]等最早提出细胞内源性ATP的含量可以反映细胞的活性和活细胞的数量。同年Gronroos[2]等也证实该技术是一种可靠、灵敏度高的确定细胞活性度的检测方法。20世纪80年代，英国人首先研制出ATP检测仪检测系统，随后发展到欧洲、美国和日本。应用范围涉及食品加工、超市和饮食行业，检测内容包括微生物和食品残渣。1998年，日本国会颁布了《关于食品制造过程管理高度化临时措施法》，其中即包含了应用ATP检测仪检测系统的内容。1999年，日本还成立了ATP涂抹检查研究会，专门研究该方法的使用效率和应用领域，其内容之一就是在食品卫生监测领域中，解决现场微生物的检测问题。20世纪末，一些ATP检测仪检测系统及技术被引进我国，到目前为止，除个别省级卫生监督检测单位装备外，主要是在一些外资或合资企业中自行检测使用。2002年，我国卫生部颁发了食品加工企业HACCP实施指南，鼓励食品加工企业引入ATP检测系统。近年来，虫荧光素酶已通过基因工程生产，价格大幅度降低，随着相关仪器的小型化，ATP生物发光技术必将会在国内相关行业得到迅速普及[3] 。 2 ATP生物发光法菌落计数与传统方法的比较 2.1 ATP生物发光法菌落计数 ATP广泛存在于各种活的生物体中，活的菌体中也含有ATP。细菌死亡后，在细胞内酶的作用下，将很快被分解掉。ATP生物荧光检测是基于生物发光体系的发光机理所设计，萤火虫具有特殊的发光物质——虫荧光素及荧光素酶，荧光素易被氧化
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如何从粪便中快速提取基因组DNA（无抑制物，可直接做PCR）
作者：德尔塔 日期：2022-05-04
如何从粪便中快速提取基因组DNA（无抑制物，可直接做PCR） 关键词：粪便DNA提取 粪便基因组提取 基因组DNA提取 北京艾德莱提供粪便基因组DNA快速提取采用硅胶膜技术从新鲜或冷冻人类粪便或其他样品类型（混有高浓度的PCR抑制剂）中抽提多至30μg 的基因组、细菌、病毒和寄生物DNA。独特的吸附树脂配合经优化的缓冲液可去除PCR抑制剂。方便的Aidlab 离心柱纯化过程只需40分钟，用于从粪便中纯化最多30μg 基因组、细菌、病毒和寄生虫DNA；快速纯化高品质，即用型DNA；无有机提取或乙醇沉淀；持续的高产量；彻底去除污染物和下游反应抑制物。纯化的DNA长度可达50 kb。该长度的DNA变性完全，并具有最高的扩增效率。高纯度的DNA可直接用于下游扩增反应。原理，方法，操作一致。完美替代Qiagen公司QiaAmp DNA Stool Mini Kit。 详细介绍： v 产品介绍： 常规的DNA纯化方式并不能有效地去除粪便中存在的大量抑制因子而导致下游实验的失败，如PCR不能扩增出所需片段。该试剂盒采用DNA吸附柱和新型独特的溶液系统，能有效去除动物粪便中各种影响下游实验（如PCR）的抑制因子，并能高效地回收粪便中的基因组DNA。动物粪便样品经特殊缓冲液ASL重悬后，70℃处理5分钟裂解细菌；离心去除不溶解的杂质，蛋白酶K消化进一步去除蛋白和杂质；然后基因组DNA在高离序盐状态下选择性吸附于离心柱内硅基质膜， 再通过一系列快速的漂洗－离心的步骤， 抑制物去除液和漂洗液将细胞代谢物、蛋白等杂质去除， 最后低盐的洗脱缓冲液将纯净基因组DNA从硅基质膜上洗脱。 v 产品特点： 1. 离心吸附柱内硅基质膜全部采用进口世界著名公司特制吸附膜，柱与柱之间吸附量差异极小，可重复性好。克服了国产试剂盒膜质量不稳定的弊端。 2. 不需要使用有毒的苯酚等试剂，也不需要乙醇沉淀等步骤。 3. 快速，简捷，单个样品操作一般可在40分钟内完成。 4. 多次柱漂洗确保高纯度，OD260/OD28
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