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研究技术支持机构(rtsf) 在365bet体育的支持和对研究和研究生课程的副院长办公室的主持下进行管理。

在rtsf内核提供了在流式细胞仪,基因组学,质谱学和蛋白质组学365bet体育的生物医学和农业研究技术和分析支持。流式细胞术芯的流动提供了两个多的基于激光的光学流式细胞仪,其同时分析细胞群为多个特性。基因组学的核心提供单基因与基因组规模DNA测序服务,片段分析基因分型,QPCR和其他核酸QC服务。蛋白质组学芯提供高通量质谱的蛋白质鉴定和的蛋白表达模式判定。质谱核使用的小分子的识别,以及分析和代谢组学分析质谱技术的阵列。

 

流式细胞术芯

This facility has available two flow cytometers – an Influx cell sorter and an LSR II for cell analysis only. The Influx is capable of sorting cells from bacteria to plant protoplasts in size from low to high speeds as dictated by the cells being sorted. Single to 6 way collection of sorted cells is available including collection to a variety of culture plates and tubes. The Influx is a BSL2+ capable sorter installed in a Baker hood with the Influx having additional aerosol collection safety equipment. 该 machine is equipped with 5 lasers. A 561 laser is used to excite PE and its conjugates (PE, PE-Texas Red, PE-Cy5 or Cy5.5, and PE-Cy7) as well as mCherry, mTomato, and others. A blue laser (488nm) is used to excite FITC, PI, PerCP-Cy5.5 with a filter set for GFP/YFP. A red laser (640 nm) is used to excite APC, Alexa 700 and APCCy7. A violet laser (405nm) is used to excite cyan proteins or Pacific Blue, and Pacific Orange. A uv laser (355nm) allows use of indo-1 calcium flux, Hoechst 342 red/blue stem cell side population and DAPI for cell cycle analysis. Data acquisition is with Sortware.

在LSR II配有3个激光器。蓝色激光具有4个检测器能够检测FITC,PE,PE-CY7和下面的(PE-txred,PE-Cy5的,percpcy5.5)中的一个。它有一个红色激光具有三个检测器,用于660纳米,700纳米和780纳米的荧光。它也有一个紫色激光(405)与两个检测器,其是用于紫色激发青色蛋白或太平洋蓝和太平洋橙有用。数据采集​​是耍大牌的软件。两台机器使用fcs3.0数据文件格式导出到收购后的分析。

软件进行数据分析可用于细胞周期分析(MODFIT),和表型(winlist 6和的FlowJo)。种类繁多的植物和动物细胞的研究方法可以在这些机器上被执行。凋亡性细胞死亡,细胞增殖,细胞活化和信号传导,细胞完整性,细胞和细胞器的功能,和存在或不存在等离子体膜,细胞质或细胞核标记物多达9个或15的标记可以同时分别进行,LSR II或流入。细胞DNA的研究可以做确定非整倍体和多倍体的DNA含量和存在或不存在。

 

基因组学核心

The MSU 基因组学核心 provides small scale to genome size high throughput sequencing services. 该 Core also provides instrumentation for SNP profiling, fragment analysis genotyping, Quantitative Real Time PCR and other nucleic acid QC services. Equipment within the Core includes an Illumina HiSeq 4000, an Illumina NextSeq 500 and two Illumina MiSeqs for next-generation sequencing. An Oxford Nanopore GridION is available for long-read sequencing, and an Oxford Nanopore PromethION will be introduced to the Core in the summer of 2019. An ABI 3730xl 96-capillary DNA sequencer performs traditional Sanger sequencing, fragment analysis and SNP sequencing. A TakaraBio SmartChip RT-PCR System and an Applied Biosystems QuantStudio 7 Flex are used for qPCR analysis. Gene expression analysis may also be performed on a NanoString nCounter. A Biomek FX robot and a BioMek 3000 robot are available for repetitive liquid handling tasks. Size-specific shearing of nucleic acids is achieved with a Covaris M220 Sonicator. An Agilent 4200 TapeStation and an Agilent 2100 Bioanalyzer are used for RNA and DNA quality and size analysis. Fluorescence-based nucleic acid quantification is performed on either an Invitrogen Qubit or a FLUOstar OPTIMA plate reader.

 

质谱核心

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蛋白质组学核心

蛋白质组学芯具有在它的现有技术的LC / MS / MS质谱仪的处置2状态;在Thermo Scientific Q-exactive和在该rmo Scientific线性离子阱(LTQ)。所述q exactive通过接口与一个热容易NLC-1000 UPLC和挠曲喷雾电离源。一起本系统结合独特的高分辨率/精确质量测量能力和高灵敏度检测的Orbitrap具有卓越质量选择和过扩展,稳定LC梯度频谱复用从而非常适用于复杂样品的蛋白质组深剖析和高度特异性的肽定量。的LTQ连接到水域纳米ACQUITY UPLC,并提供高灵敏度,快速扫描速度和毫秒n 具有卓越的色谱分离沿能力。该设施提供了使用吉祥物蒸馏,蛋白质组发现者,maxquant和天际线软件套件的数据处理和定量。吉祥物,SEQUEST和仙女座搜索算法可用于肽与频谱匹配以及支架用于数据可视化和统计验证。前面复合肽样品的色谱分离时使用Waters ACQUITY h级UPLC和Agilent 3100 OFFGEL分馏器提供。可以使用Invitrogen变焦IEF精馏塔或用1d和2d页进行蛋白水平的分离。