Recombinant Human His6-GABARAP Protein, CF Summary
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
UL-410
| Formulation | X mg/ml (X μM) in 50 mM HEPES pH 8.0, 100 mM NaCl, 10% (v/v) Glycerol |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Reconstitution Calculator
Background: GABARAP
gamma -Aminobutyric Acid Receptor-associated Protein (GABARAP) is an 117 amino acid (aa) protein with a predicted molecular weight of 14 kDa. It is a member of the GABARAP subfamily of the Autophagy-related 8 (ATG8) family of proteins (1). The GABARAP subfamily also includes GABARAPL1. GABARAP has 100% aa sequence identity with its mouse and rat orthologs and is orthologous to yeast ATG8. ATG8 family members show structural similarity with Ubiquitin, but lack aa sequence similarity. GABARAP was first described for its putative involvement GABA A Receptor trafficking (2,3). However, it is best known for its role in autophagy (1,4). GABARAP covalently attaches to phosphatidylethanolamine (PE) in the phagophore (autophagosome precursor) membrane using a Ubiquitin-like conjugation system that includes Ubiquitin-activating (E1)-, Ubiquitin-conjugating (E2)-, and Ubiquitin Ligase (E3)-like enzymes. Here it is involved in the later stages of autophagosome maturation (4,5). It may also be involved in cargo recruitment to autophagosomes (1).
- Shpilka, T. et al. (2011) Genome Biol. 12:226.
- Wang, H. et al. (1999) Nature 397:69.
- Leil, T.A. et al. (2004) J. Neurosci. 24:11429.
- Weidberg, H. et al. (2010) EMBO J. 29:1792.
- Weidberg, H. et al. (2011) Ann. Rev. Biochem. 80:125.
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2 单细胞测序是指单个细胞水平上对基因组进行测序。
3、靶向测序步骤为 样品准备、探针/引物设计、目标序列捕获、文库制备、上机测序、数据分析。
4、单细胞的分离--DNA/RNA的提取和扩增（全基因组扩增和全转录组扩增）---测序以及后续的分析和应用。
相同:使用二代测序
不同：样本处理，文库构建，检测的区域，数据分析流程
目前，市面上质粒提取试剂盒大多数都是采取上述的碱裂解方法，不同之处在于纯化方式。目前比较常用的纯化系统是硅基质吸附材料，其原理为在高盐环境下质粒DNA能够结合到硅基质上，然后用低盐缓冲液或水将质粒DNA从硅基质上洗脱下来。得到的质粒可以用于酶切、PCR、测序、细菌转化、转染等分子生物学实验。
2．pgem-3zf (+) 双链dna对照模板 0.2g/l，试剂盒配套试剂。
3．m13(-21)引物 tgtaaaacgacggccagt，3.2μmol/l，即3.2pmol/μl，试剂盒配套试剂。
4．dna测序模板 可以是pcr产物、单链dna和质粒dna等。模板浓度应调整在pcr反应时取量1μl为宜。本实验测定的质粒dna，浓度为0.2g/l，即200ng/μl。
5．引物 需根据所要测定的dna片段设计正向或反向引物，配制成3.2μmol/l，即3.2pmol/μl。如重组质粒中含通用引物序列也可用通用引物，如m13(-21)引物，t7引物等。
6．灭菌去离子水或三蒸水。
7．0.2ml或和0.5ml的pcr管 盖体分离，pe公司产品。
8．3mol/l 醋酸钠(ph5.2) 称取40.8g naac·3h2o溶于70ml蒸馏水中，冰醋酸调ph至5.2，定容至100ml，高压灭菌后分装。
9．70%乙醇和无水乙醇。
10．naac/乙醇混合液 取37.5ml无水乙醇和2.5ml 3mol/l naac混匀，室温可保存1年。
11．pop 6测序胶 abi产品。
12．模板抑制试剂(tsr) abi产品。
13．10×电泳缓冲液 abi产品。
14．abi prism 310型全自动dna测序仪。
15．2400型或9600型pcr仪。
16．台式冷冻高速离心机。
17．台式高速离心机或袖珍离心机。向左转|向右转
亲子鉴定？
ID,promega.我们实验室主要用的是这两种，由于怕有突变还会备有凯杰的。
如果测序的话，那么就是AB的bigdye吧？
有什么问题我们可以交流。
数据分析的小白一枚，最近开始学习二代测序仪IONTORRENT出来的靶向基因检测位点的数据分析。由于完全没有涉猎过这个领域，会有很多不懂得地方，望大神们不吝赐教！
比如上面这个搜索结果，就是根据一起出来的数据，也就是一个位点的rs号进行检索出来的界面，但是完全不知道如何下手，也不是很清楚具体模块的作用，我能得到什么结果。虽然在网上能看到一些说明，但是仍然一知半解的！
最后，还是想说，大神们快快带我入门吧，非常迫切！跪谢！
