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核酸纯化

核酸纯化柱

分子生物学试剂

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全血DNA小量试剂盒 创新

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产品序号:3001050 / 3001250

规格价格
50次制备520
250次制备2350

适合在15~20分钟内从全血中快速分离纯化基因组DNA

全血标本用量:200~400μl

无需事先分离去除红细胞

无需蛋白酶K消化步骤



适用于从全血中分离纯化最多达15 μgDNA

·15分钟内即可完成血液总DNA的制备。
·
无须事先分离去除红细胞及蛋白酶K消化步骤,可室温运输储存。
·
可从新鲜或者是冷冻的全血、溶血的全血、唾液、 细胞悬浮液等标本中分离纯化总DNA。       
·
核酸纯化柱可最大吸附15μg DNA
·
起始样品体积:200 μl- 400 μl全血。
·
彻底清除血样中的PCR抑制物,可使用多至1/2反应体系体积的模板进行扩增。
·
所需仪器:可适合2 ml离心管使用的离心机。

产品原理
全血样品经裂解液溶解后,用沉淀液沉淀血红蛋白。柱纯化核酸技术过滤除去残留的血红蛋白,吸附在纯化柱上的总DNABuffer WABuffer WB洗涤后,可彻底清除残留在纯化柱上的杂质及PCR抑制物。纯化柱上的总DNA直接用BufferTE或水洗脱,并可立即用于各种分子生物学实验。



操作视频


操作步骤
400 μl全血中加入300 μl Buffer L1溶解全血释放DNA,再加入300μl Buff

er L2沉淀血红蛋白,离心取上清加入纯化柱中,经过结合、清洗步骤去除残留的杂质后,DNA即可被TE溶液洗脱下来。

可适用的分子生物学实验


1PCR
2
RFLP分析
3
Southern blotting
4.
基因分型,包括指纹印迹和亲子鉴定

常见问题分析

1、回收不到DNA或者DNA的回收效率低

可能的原因:

1)Buffer WA Buffer WB中未加入无水乙醇,应按比例补加无水乙醇。如果是错误地加入了其他试剂,请向我公司技术部寻求帮助。

2)Buffer WA Buffer WB中错误地加入了70%乙醇。请向我公司技术部寻求帮助。

3)错误地使用了Buffer WA Buffer WB的洗涤顺序。确保按正确的顺序洗涤纯化柱。

4)全血样品保存不当,导致全血中的DNA降解。在2-8℃冰箱放置超过两个星期的全血标本开始出现溶血现象,如果需要继续使用血样,则应转入-20℃-70℃冻存。否则获得的DNA将开始出现凋亡细胞的DNA带型,并且此时全血中的DNA随着时间的延长会持续分解,直至分离不到电泳可见的DNA

5)DNA的洗脱效率差。参考第4页柱纯化技术中的第4点“洗脱DNA“内容优化DNA的洗脱方案。

6)错误地使用了Buffer L1 Buffer L2的溶解顺序。确保按正确的顺序操作。

 

2.       纯化柱膜上残留有血色素

 可能的原因:

Buffer WA Buffer WB中未加入无水乙醇,应按比例补加无水乙醇。

 

3.       样品间交叉污染

避免样品间交叉污染的方法:

1) 选用含有滤芯的移液器吸头用于实验的操作。

2) 实验操作时勿使吸头触碰到纯化柱。

3) 丢弃滤液后将2ml离心管在纸巾上倒扣拍击一次,以避免粘附在离心管管口的滤液对离心机的污染。

4) 如果手套接触到标本,应及时更换手套。


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1. Zhu Y, Shentu X, Wang W, et al. A Chinese family with progressive childhood cataracts and IVS3+ 1G> A CRYBA3/A1 mutations[J]. Molecular vision, 2010, 16: 2347.   影响因子:1.986

 

2. 四川地区猕猴线粒体DNA?控制区遗传多样性及其种群遗传结构

徐怀亮, 李地艳, 程安春, 姚永芳, 倪庆永, 曾文… - 兽类学报, 2010 - mammal.cn

 

3.  ABCB1 基因多态性与母胎界面糖皮质激素的转运关系

李健, 王自能 - 暨南大学学报(自然科学与医学版), 2011 - jnxb.jnu.edu.cn

 

4. The TGFBI R555W mutation induces a new granular corneal dystrophy type I phenotype

Y Zhu, X Shentu, W Wang - Molecular vision, 2011 - ncbi.nlm.nih.gov

 

5. 藏酋猴Mhc-DPB1 基因exon 2 的多态性

李佳薏, 姚永芳, 周亮, 徐怀亮 - 遗传, 2012 - chinagene.cn

 

6. Molecular analysis of SLC25A13 gene in human peripheral blood lymphocytes: Marked transcript diversity, and the feasibility of cDNA cloning as a diagnostic tool for …

ZH Zhang, WX Lin, M Deng, XJ Zhao, YZ Song - Gene, 2012 - Elsevier

 

7. Association of fetal but not maternal P-glycoprotein C3435T polymorphism with fetal growth and birth weight, a possible risk factor for cardiovascular diseases in …

J Li, ZN Wang, YP Chen, YP Dong, XM Xiao… - Clinical …, 2012 - uni-potsdam.de

 

8. A novel p. R890C mutation in EPHA2 gene associated with progressive childhood posterior cataract in a Chinese family

XC Shentu, SJ Zhao, L Zhang… - International journal of …, 2013 - ncbi.nlm.nih.gov

 

9. Novel mutation c. 980_983delATTA compound with c. 986C> A mutation of the FRMD7 gene in a Chinese family with X-linked idiopathic congenital nystagmus

F Song, B Chen, Z Sun, L Wu, S Zhao, Q Miao… - Journal of Zhejiang …, 2013 - Springer

 

10. Chinese family with atypical granular corneal dystrophy type I caused by the typical R555W mutation in TGFBI

SJ Zhao, YN Zhu, XC Shentu, Q Miao - International journal of …, 2013 - ncbi.nlm.nih.gov

 

11.Characterization of the Relationship between APOBEC3B Deletion and ACE Alu Insertion

K Wang, Y Li, C Dai, K Wang, J Yu, Y Tan, W Zhang… - 2013 - dx.plos.org

 

12. A functional fetal HSD11B2 [CA] n microsatellite polymorphism is associated with maternal serum cortisol concentrations in pregnant women

J LiYP ChenZN WangTB LiuD Chen… - Kidney and Blood …, 2013 - karger.com

 

13. A missense mutation in the arginine‐vasopressin neurophysin‐II gene causes autosomal dominant neurohypophyseal diabetes insipidus in a Chinese family

D Ye, FQ Dong, WQ Lu, Z Zhang, XL Lu… - Clinical …, 2013 - Wiley Online Library

 

14. Molecular diagnosis of a Chinese pedigree with α-mannosidosis and identification of a novel missense mutation

X Wu, J Pan, Y Guo, C Guo, W Jiang, R Li… - Journal of Pediatric …, 2014 - degruyter.com

 

15. A novel MIP gene mutation associated with autosomal dominant congenital cataracts in a Chinese family

Y Yu, Y Yu, P Chen, J Li, Y Zhu, Y Zhai… - BMC medical …, 2014 - biomedcentral.com

 

16. Molecular analysis and phenotypic study in 14 Chinese families with Bietti crystalline dystrophy

H Yin, C Jin, X Fang, Q Miao, Y Zhao, Z Chen, Z Su… - PloS one, 2014 - dx.plos.org

 

17. Identification of a novel frameshift mutation in PITX2 gene in a Chinese family with Axenfeld-Rieger syndrome

H Yin, X Fang, C Jin, J Yin, J Li, S Zhao, Q Miao… - Journal of Zhejiang …, 2014 - Springer

 

18. CFI-rs7356506 is a genetic protective factor for acute anterior uveitis in Chinese patients

Y Wang, XF Huang, MM Yang, WJ Cai… - British Journal of …, 2014 - bjo.bmj.com

 

19. Identification and Functional Analysis of a Novel MIP Gene Mutation Associated with Congenital Cataract in a Chinese Family

X Shentu, Q Miao, X Tang, H Yin, Y Zhao - 2015 - dx.plos.org

 

20. Association of CD40-1C/T Polymorphism in the 5′-Untranslated Region with Chronic HBV Infection

C Zhou, X Jin, J Tang, J Fei, C Gu… - Cellular Physiology and …, 2015 - karger.com

 

21. A novel phenotype-genotype correlation with an Arg555Trp mutation of TGFBI gene in Thiel-Behnke corneal dystrophy in a Chinese pedigree

Y Yu, P Qiu, Y Zhu, J Li, M Wu, B Zhang… - BMC …, 2015 - biomedcentral.com

 

22.CFI-rs7356506 polymorphisms associated with Vogt-Koyanagi-Harada syndrome

ML Dai, XF HuangQF Wang, WJ Cai, ZB Jin… - Molecular …, 2016 - ncbi.nlm.nih.gov

 

23.Association of polymorphisms of IRF6 to non-syndromic cleft lip with or without palate in a Guangdong population

W Wu, J Hao, H Wang, L Hua, F Li, Y Chen… - … JOURNAL OF CLINICAL …, 2016 - ijcem.com

 

24.SLC25A13 cDNA cloning analysis using peripheral blood lymphocytes facilitates the identification of a large deletion mutation: Molecular diagnosis of an infant with …

HS Zeng, WX Lin, ST Zhao… - Molecular …, 2016 - spandidos-publications.com

 

25.Molecular Epidemiological Survey of Glucose-6-Phosphate Dehydrogenase Deficiency and Thalassemia in Uygur and Kazak Ethnic Groups in Xinjiang, Northwest …

L Han, H Su, H Wu, W Jiang, S Chen - Hemoglobin, 2016 - Taylor & Francis

 

26.CFHR2‐rs2986127 as a genetic protective marker for acute anterior uveitis in Chinese patients

XF HuangY Wang, FF Li, D Lin, ML Dai… - The journal of gene …, 2016 - Wiley Online Library

 

27.Association of CD59 and CFH polymorphisms with acute anterior uveitis in Chinese population

QF WangXF Huang, ZL Zheng, ML Dai, WJ Cai… - Eye, 2016 - nature.com

 

28.A novel splice site mutation of CRYBA3/A1 gene associated with congenital cataract in a Chinese family

MH Wu, YH Yu, QL Hao, XH Gong… - International Journal of …, 2017 - ncbi.nlm.nih.gov

 

29.Genome-Wide Detection of Copy Number Variations in Unsolved Inherited Retinal Disease Genome-Wide Detection of CNVs in Unsolved IRDs

XF Huang, JY Mao, ZQ Huang, FQ Rao… - … & Visual Science, 2017 - iovs.arvojournals.org

 

 30. Citrin缺陷导致的新生儿肝内胆汁淤积症患儿SLC25A13基因突变与生化指标的相关性研究

 

31. Citrin缺陷病致病基因SLC25A13高频突变筛查试剂盒

   宋元宗张占会 

 


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