一般描述
GenElute血液基因组DNA试剂盒可以简单、方便地从新鲜或长时间存放（超过24小时）的全血中分离高纯度基因组DNA。该试剂盒将硅胶膜技术的优势与微量离心形式结合在一起，同时无需采用昂贵的树脂、乙醇沉淀，以及诸如苯酚和氯仿等有害化学成分。
应用
GenElute™血液基因组DNA试剂盒已被用于：
- 从切除输精管的兔子体内获得的大颗粒（LG）和小颗粒（SG）样品中提取DNA
- 从血液样本中分离DNA
- 从剩余血细胞中提取全血DNA
纯化所得的基因组DNA可用于以下下游应用：
- 限制性内切酶酶切
- PCR
- Southern印迹
- 测序反应
- 克隆
特点和优势
- 起始材料：不超过200 μl新鲜或长时间存放的血液
- 预期得率：至多10 μg
- 洗脱体积：400 μl
- 所需时间：<40分钟
- A260/A280比率：1.6 - 1.9
- 兼容多种抗凝血素、包括EDTA、肝素和柠檬酸钠
原理
起始材料首先溶解在含有离液盐的溶液之中，以确保大分子彻底变性。之后加入乙醇，从而使DNA在溶解物通过微量离心管在硅胶膜上离心时，结合到膜上。随附一份Prewash Solution，可帮助与全血样本长时间存放（超过24小时）相关的污染物。在洗涤去除污染物后，将DNA放在200 mL的Tris-EDTA溶液中洗脱。
预期基因组DNA得率取决于所用的起始材料的数量和性质（例如，从200 μl 新鲜的全血之中，可在一小时内分离出4至10 μg RNase A处理的DNA）。通过本试剂盒纯化所得的DNA的A260/A280比率介于1.6-1.9之间，最大长度可达50 kb。
预期基因组DNA得率取决于所用的起始材料的数量和性质（例如，从200 μl 新鲜的全血之中，可在一小时内分离出4至10 μg RNase A处理的DNA）。通过本试剂盒纯化所得的DNA的A260/A280比率介于1.6-1.9之间，最大长度可达50 kb。
其他说明
更多信息，请见www.sigma-aldrich.com/genomicdna。
法律信息
GenElute is a trademark of Sigma-Aldrich Co. LLC
警示用语：
Danger
危险分类
Acute Tox. 4 Oral - Aquatic Acute 1 - Aquatic Chronic 2 - Eye Dam. 1 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
靶器官
Respiratory system
储存分类代码
10 - Combustible liquids
法规信息
常规特殊物品
Yanxia Gao et al.
Renal failure, 36(8), 1226-1232 (2014-07-01)
The objective of this study is to identify ATP6V1B1, ATP6V0A4 and SLC4A1 genes mutations and assess audiologic characteristics in six Chinese children with primary distal renal tubular acidosis from four unrelated families between the ages of 2 and 13 years.
Xiangzhong Zhao et al.
Scientific reports, 6, 33920-33920 (2016-09-27)
Familial renal glycosuria (FRG) is caused by mutations in the SLC5A2 gene, which codes for Na+-glucose co-transporters 2 (SGLT2). The aim of this study was to analyze and identify the mutations in 16 patients from 8 families with FRG. All
Cui Wang et al.
Scientific reports, 6, 33652-33652 (2016-09-21)
Primary hyperoxaluria type 1 (PH1) is a rare genetic disease characterized by excessive oxalate accumulation in plasma and urine, resulting in various phenotypes because of allelic and clinical heterogeneity. This study aimed to detect disease-associated genetic mutations in three PH1
Sai Wang et al.
Frontiers in genetics, 11, 585064-585064 (2020-12-01)
Familial renal glucosuria is a rare renal tubular disorder caused by SLC5A2 gene variants. Most of them are exonic variants and have been classified as missense variants. However, there is growing evidence that some of these variants can be detrimental
Involvement of macrophage migration inhibitory factor in the pathogenesis of idiopathic orbital inflammatory pseudotumor
Zhao P, et al.
International Journal of Clinical and Experimental Pathology, 9(7), 6659-6671 (2016)
实验方案
Whole genome amplification (WGA) of plasma and serum DNA presents a unique challenge due to the small amount of nucleic acid in such samples.
Protocol extracts genomic DNA from blood cards, useful for limited DNA samples in amplification.
Simple DNA isolation protocol from whole blood samples for genomic analysis, followed by GenomePlex® amplification.
The GenElute™ Blood Genomic DNA Kit Protocol provides a simple and convenient way to isolate pure genomic DNA from fresh or aged whole blood.
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