The Refolding CA Kit takes advantage of a novel artificial chaperone technology in an easy-to-use two-step process to optimize refolding conditions of inclusion body proteins. Optimization of protein refolding conditions results in correct protein folding and restoration of protein activity. One can also use this kit to solubilize inclusion body proteins.
The Refolding CA (Cycloamylose) Kittakes advantage ofa novel artificial chaperone technologyin an easy-to-use two-step process to optimize refolding conditions of inclusion body proteins. Optimization of proteinrefoldingconditions results in correct protein foldingand restoration of protein activity. One can also use this kit to solubilize inclusion body proteins.
The smaller-size Refolding CA Kit is supplied with guanidine hydrochloride (GdmCl) and dithiothreitol (DTT) for protein unfolding, four different surfactants that can be added independently to the unfolded protein solution (protecting the proteinfrom molecular aggregation), and highly polymerized cycloamylose (CA), an artificial chaperone, for surfactant removal and recovery of protein activity. Overnight incubation of the CA-treated protein is followed by a quick 10-min centrifugation step and collection of the refolded protein-containing supernatant.
The Refolding CA Large Kit isintended for large-scale protein refolding after reaction conditions have been optimized using the smaller Refolding CA Kit. The large kit consists of only denaturant and CA.
The artificial chaperone technology in the Refolding CA Kit is licensed from NFRI, BTRAI and Ezaki Glico Co, Ltd.
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逆转录（reverse transcription）是以RNA为模板合成DNA的过程，即RNA指导下的DNA合成。此过程中，核酸合成与转录（DNA到RNA）过程与遗传信息的流动方向（RNA到DNA）相反，故称为逆转录。逆转录过程是RNA病毒的复制形式之一，需逆转录酶的催化。 逆转录过程的揭示是分子生物学研究中的重大发现，是对中心法则的重要修正和补充。人们通过体外模拟该过程，以样本中提取的mRNA为模板，在逆转录酶的作用下，合成出互补的cDNA，构建cDNA文库，并从中筛选特异的目的基因。该方法已成为基因工程技术中最常用的获得目的基因的策略之一。
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。
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大家miRNA逆转录和qPCR的试剂盒用的是那个公司的啊，求推荐！
