
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)已包被抗原或抗体的固相载体(免疫吸附剂);
(2)酶标记的抗原或抗体(结合物);
(3)酶的底物;
(4)阴性对照品和阳性对照品(定性测定中),参考标准品和控制血清(定量测定中);
(5)结合物及标本的稀释液;
(6)洗涤液,在板式ELISA中,常用的稀释液为含0.05%吐温20磷酸缓冲盐水;
(7)酶反应终止液,常用的HRP反应终止液为硫酸,其浓度按加量及比色液的最终体积而异,在板式ELISA中一般采用3mol/L。向左转|向右转
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。
现在跑tricine-sds-page电泳老是跑不好,
我刚做了bradford定量,
觉得差异与空菌对照还是挺明显的,
但是这个方法只能初略的估计,
还是不能精确定量,
做HPLC我现在还没有条件,
所以我想能不能用BCA或者lowry法先进行定量啊?
这种方法如果做出来结果的话,
能不能发个sci之类的文章啊?
希望高手们指点一下!
谢谢了
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。

