一、Agilent表达谱芯片
基因表达谱芯片是最早实现从高通量的角度检测全基因组差异基因表达的平台。为方便广大科研工作者转录组基因表达筛选分析,中科普瑞引进Agilent表达谱芯片检测平台,为您的转录组mRNA检测和分析提供全方位的服务。在美国FDA组织的MAQC中,作为众多参与该计划的芯片制造商之一,Agilent的表达谱芯片产品无论从检出率,芯片的跨平台比较,数据重复性,平均CV,还是与qPCR的一致性等方面,均表现出卓越的性能。
² 独特的喷墨原位合成技术;
² 探针长度60 mer,同时兼顾较高的灵敏性和特异性;
² 由于探针设计较长,芯片检出率高,有利于捕捉对生命活动有更重要意义的低丰度基因;
² 芯片随信息库及时更新,基因信息更新更全,可不断升级到高级芯片版本;
² 灵活的定制方案,客户可任意制作自己的芯片,仅需基因组或EST序列;
² 平均CV<10% (MAQC: Nature Biotechnology Sept, 2006);
² 芯片具有良好的可重复性,相关系数R2>0.95 (MAQC);
² 芯片数据结果与qPCR数据结果具有良好的相关性,相关系数R2>0.9 (MAQC)。
拟南芥、玉米、水稻、烟草、牛、鸡、狗、马、猪、羊、兔、猴、蛙、三文鱼、斑马鱼、线虫、蚊子、果蝇、酵母、大肠杆菌、稻瘟菌(更多物种或需要定制请咨询销售人员)
1. 样品纯度:OD 260/280值应在1.9~2.2 之间;DNA 应该去除干净;
2. 样本完整性:Agilent 2100 RIN≥ 7,且28S/18S≥ 0.7;
3. 样品浓度:低浓度不低于100ng/µl;
4. 样品总量:每个样品总量不少于2µg;
5. 样品溶剂:要溶解在RNase-free H20或RNase-free TE (pH 8.0)中;
6. 样品运输:RNA用冻存管保存,并用干冰或液氮运输。
基础分析 | |
1. 原始数据处理分析 | 1)数据归一化 |
2)箱线图 | |
3)样本聚类图 | |
4)样本相关性分析 | |
5)主成份分析 (PCA) | |
2. 差异基因筛选 | 1)表达差异基因筛选 |
2)散点图 | |
3)火山图 | |
4)差异基因层级聚类热图 | |
3. 差异mRNA功能分析(GO/KEGG) |
1. 原始数据处理分析
2. 差异基因筛选
3. 差异mRNA功能分析(GO/KEGG)
4. 信号通路调控网络构建(Pathway-Network)
信号通路调控网络构建(Pathway-Network)是根据所有差异基因同时参与的Pathway之间的相互调控关系构建信号通路调控网络,从系统的角度研究各个信号通路间的信号传导和调控过程,在多个显著性Pathway中发现受实验影响的核心Pathway,以及实验影响的信号通路之间的调控机理.
Pathway-Network
5. 基因相互作用网络图
基因网络是一组基因的集合体,这些基因通过相互协作来控制生物体重要的生命过程。我们通过筛选的差异基因进行基于实验验证,其他亲缘物种数据库检索、文献挖掘等多种信息进行基因互作网络的构建。
基因相互作用网络图
6. 转录因子分析及网络构建
转录因子与基因表达调控区域的特异性结合是基因表达的重要调控方式。针对特定转录因子的实验,可对差异基因进行转录因子结合位点的motif分析,从而推测差异基因中有哪些基因可能直接受该转录因子调节。最后结果会用统计学方法进行检验,找到调控目标生物性状,统计学上有显著差别的转录因子。找到差异的转录因子及其调控的靶基因,我们可以构建TF-network,从中得出关键的转录因子及被关键转录因子转录调控的基因,系统的研究转录因子与基因的调控关系。
转录因子网络构建
7. 共表达网络构建(Coexpression Network)
共表达网络图(Coexpression Network)是根据基因表达信号值的动态变化,计算基因间的共表达关系,得到基因间的表达调控关系及调控方向,从而构建基因的表达调控网络。利用共表达网络图,研究者可通过分析基因调控能力,获得样本随实验变化的核心调控基因。
要求:每组数据3个或3个以上生物学重复
8.时间序列的共表达网络
根据基因表达信号值随着时间序列的变化,计算基因间的共表达关系,得到基因间的表达调控关系及调控方向,从而构建基因的表达调控网络。利用此网络图,研究者可通过分析基因调控能力,获得样本在时间过程中起调控作用的基因。
时间序列的共表达网络
9.分子建模预测(MAKER 预测)
采用模式识别与数据挖掘技术有效进行预测模型的构建,将部分芯片数据拿来做预测模型,然后部分芯片数据作为测试数据集(独立样本)来验证模型的准确性。目的在于利用实验数据来筛选出一批靶标基因,并以此构建模型,以进行早期诊断、疾病预测和预后分析。
MAKER 预测
二、中科普瑞基因芯片服务平台
高标准 — 拥有15年以上科技服务实验和分析技术经验的团队成员以GLP-like质量管理体系为您的基因芯片服务体系全线保驾护航。
高效率 — 样品质检合格后,15个工作日出具结果报告,专属生物信息和技术支持服务人员为您提供在线结果咨询和解释服务。
高质量 — 符合3大基因芯片厂商质量要求的服务质量体系,项目质量管理专员负责对您的项目全程跟踪和质控。
三、中科普瑞科技服务团队参与Agilent表达谱芯片发表相关文献
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