The SMART-Seq Single Cell Kit (SSsc) and the SMART-Seq Single Cell PLUS Kit (SSsc PLUS) are powered by robust chemistry that provides unparalleled sensitivity and reproducibility for single-cell and nuclei applications. These kits use oligo(dT) priming to generate high-quality, full-length cDNA directly from single cells known to have low RNA content (e.g., PBMCs, T-cells, B-cells, etc.).
The SMART-Seq Single Cell Kit (SSsc) and the SMART-Seq Single Cell PLUS Kit (SSsc PLUS) are powered by robust chemistry that provides unparalleled sensitivity and reproducibility for single-cell and nuclei applications. These kits use oligo(dT) priming to generate high-quality, full-length cDNA directly from single cells known to have low RNA content (e.g., PBMCs, T-cells, B-cells, etc.). In addition to the cDNA synthesis kit, the SSsc PLUS kit also includes a library preparation kit and a single-use unique dual index (UDI) plate to generate Illumina-compatible sequencing libraries, making it a complete end-to-end solution.
The SMART (Switching Mechanism at 5" End of RNA Template) technology powering the SMART-Seq Single Cell kit provides full-length transcript information, enabling analysis of transcript isoforms, gene fusions, point mutations, etc. The cDNA kit provides high reproducibility, even gene-body coverage, and an accurate representation of GC-rich transcripts. We have further modified this powerful core technology to create a new chemistry with higher sensitivity designed specifically for single-cell applications. These enhancements result in more useful reads and a higher gene detection, allowing for a better understanding of precious samples.
The SSsc PLUS kit includes our patented library preparation kit that incorporates enzymatic fragmentation and stem-loop adapters to construct high-quality, Illumina-compatible libraries from the cDNA synthesized with the core kit. This two-step workflow takes place in a single tube and can be completed in about two hours with no intermediate purification steps necessary—minimizing sample mix-up, sample loss, and cross-contamination. The high library yield generated from this kit provides the flexibility to sequence on high-throughput sequencers while providing high reproducibility.
If you are interested in these benefits but are working with intact, high-quality cells (<1,000) or ultra-low total RNA inputs, we recommend our SMART-Seq v4 PLUS Kit.
If you prefer a random priming approach that will allow you to work with degraded samples and includes library preparation and indexing reagents, we recommend our SMART-Seq Stranded Kit.
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定性ELISA只能粗略表示样本待测物含量在某个特定值以上或以下，定性ELISA通常设置阳性对照（P）、和阴性对照（N），结果分别用“阴性”和“阳性”表示。ELISA定性测定的“阴性”和“阳性”的判断标准是试剂盒所确定的阳性判断值，即cut-off值。
定量ELISA是通过一系列不同已知浓度的标准品所对应的OD值做出标准曲线，然后将样本的OD值代入标准曲线，计算出样本中待测物的含量。百特纯大分子Meretciel的定量ELISA试剂盒还可以。
1.步骤简单，45分钟内完成测定，比经典Lowry法快4倍而且更加方便。
2.灵敏度高，检测浓度下限达到25μg/ml，最小检测蛋白量达到0.5μg，待测样品体积为1-20μl 。
3.BCA法测定蛋白浓度不受绝大部分样品中的去污剂等化学物质的影响，可以兼容样品中高达5%的SDS，5%的Triton X-100，5%的Tween 20，60，80。
4.在20-2000μg/ml浓度范围内有良好的线性关系。
5.检测不同蛋白质分子的变异系数远小于考马斯亮蓝法蛋白定量。
1.模板提取（一般为RNA）：Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定：分光光度计或NanoDrop
3.逆转录：逆转录试剂盒（或者一步法试剂盒），这一步可以用普通PCR做，也可以用水域做。
4.荧光定量PCR试剂：通常有用SYBR Green Mix做的，但是这里建议你用EvaGreen做，灵敏度和平行性都要好于SYBR Green，并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他：除了以上的那些还需要离心管、PCR管或板（Axygen反应比较好）、移液枪等，暂时就想到这么多。
试剂盒里有详细的说明书，告诉你样品需要多少量，每个试剂需要加入多少量，和详细的实验步骤，一般买来就可以用，不用人教。
所以你问一个样需要多少量是没法回答的，测定过程是要加很多种试剂的。
加入荧光标记探针，巧妙地把核算扩增、杂交、光谱分析和实时检测技术结合在一起，借助于荧光信号来检测PCR产物。一方面提高了灵敏度，另一方面还可以做到PCR每循环一次就收集一个数据，建立实时扩增曲线，准确地确定CT值，从而根据CT值确定起始DNA的拷贝数，做到真正意义上的DNA定量。另外由于CT值是一个完全客观的参数，CT值越小，模版DNA的起始拷贝数越小。因此，利用CT值确定DNA拷贝数实时PCR方法比普通终点定量方法更加准确
