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血液线粒体DNA抽提试剂盒说明书近几年,从有报告暗示糖尿病、阿尔茨海默病和线粒体DNA(mtDNA)变异有关开始,mtDNA分析变得盛行1),2),3),4)。
可是，想要抽提高纯度mtDNA，需要繁杂的操作和很多的时间，不适合以多样品处理为目的的研究。
目前进行的mtDNA分析，从细胞中抽提total DNA时，产物中含有微量mtDNA与大量基因组DNA。
mtDNA 抽提WB试剂盒可短时间从全血中容易的抽提出高纯度mtDNA，适合多样品处理。
〔特点〕Ⅰ. 简单操作，可短时间（约90分钟）抽提出mtDNA。
Ⅱ. 可从EDTA 处理血及肝素处理血中抽提出mtDNA。
Ⅲ. 抽提出的mtDNA混入了少量高纯度基因组DNA，可用于限制性内切酶处理或DNA扩增反应。
Ⅳ. 适合少量多样品的DNA扩增反应的预处理。
Ⅴ. 不使用苯酚、氯仿等有害剧毒溶剂。
〔内容〕1) Leukocyte Isolation Solution 5.0 ml 1支2) DNA Extraction SolutionⅠ 26.5 ml 1支3) DNA Extraction SolutionⅡ(A) 1.3 ml 1支4) DNA Extraction SolutionⅡ(B) 1.3 ml 1支5) DNA Extraction SolutionⅢ 1.9 ml 1支6) Sodium Iodide Solution 7.5 ml 1支7) Washing Solution 50.0 ml 1支〔储存方法〕2～10℃保存〔除试剂盒外，实验所必备的东西〕试剂1) 异丙醇（特级，Code 166–04836）12.5 ml2) TE缓冲液（基因工程学研究，Code 316–90025）适量器具1) 1.5 ml 微管（已灭菌）2) 涡流搅拌器3) 微量高速离心机（最大12,000×g）4) 减压干燥机〔使用方法〕(1) 将100μl的人类新鲜血液放入微管中*1。
组织和毛发提取试剂盒（与DNA IQ TM系统结合使用）操作手册本说明书用于产品DC6740所有的技术文献均可从公司网站得到请访问公司网站以证实您所使用的技术手册为最新版本I．介绍 (1)III. 产品组分 (2)III．组织和毛发提取试剂盒与 DNA IQ TM系统结合使用的操作步骤 (2)A．试剂的准备 (2)B．从发囊及发根中纯化DNA (5)C．从新鲜组织、冰冻组织、甲醛固定组织及石蜡包埋组织中纯化DNA (6)IV．疑难解答 (7)V．参考文献 (7)VI．相关产品 (8)I.介绍Promega的 DNA IQ TM系统使用一种新的顺磁性树脂来纯化DNA, 该试剂盒中包含有DNA纯化所需的可用于多种形式样品的高效裂解液。
然而，DNA IQ TM 系统中的裂解液对一些如组织和毛发的样品不能有效地裂解，这些样品需要蛋白酶K的预处理以协助样品的裂解。
组织和毛发提取试剂盒（与DNA IQ TM系统结合使用）中提供的试剂和操作手册可用于大部分组织和毛发的有效裂解，除去样品中的蛋白和其它成分，随后用DNA IQ TM系统对样品中的DNA进行纯化。
基因组和线粒体DNA结合使用组织和毛发提取试剂盒（与DNA IQ TM系统结合使用）及DNA IQ TM系统，可高效地从样品中提取纯化双链或单链基因组DNA及线粒体DNA。
如果待提取的样品被微生物污染，则微生物DNA被一起分离。
可抑制PCR扩增的小于 80bp的 DNA可被选择性地去除，从而使纯化的 DNA更有效地用于 PCR扩增。
组织和毛发提取试剂盒（与DNA IQ TM系统结合使用）可提高 DNA纯化的产量当用DNA IQ TM系统—数据库操作手册（TB297）中涉及的样品进行 DNA纯化时，DNA IQ TM系统所提取的 DNA的量是固定的，约 100ng。
对于不同检材中DNA的定量提取依赖于样品中超量的DNA饱和DNA IQ TM 树脂。
在结合使用组织和毛发提取试剂盒（与DNA IQ TM系统结合使用）及 DNA IQ TM系统时，若用蛋白酶K消化去除竞争结合DNA IQ TM 树脂的物质，则树脂从蛋白酶K处理过的注意：本中文操作手册仅供实验参考，在实际使用中请详细对照原英文技术手册TB307。
PromegaDNA提取试剂盒的使用方法一、使用前的准备工作Proteinase k solution:用无核酸酶的水把蛋白酶K稀释成20mg/ml，按照自己的每次平均使用量分装，储存在-20℃，融化时要在冰上融化，反复冻融会降低Proteinase K的活性消化液配制：在tube中混合下面各种反应物，使用前放在冰上。
Wizard SV Wash solution（洗脱液）的配制：按照瓶子标签上要求的量，添加95%的乙醇到Wizard sv solution 中，做上标记，室温保存。
PromegaDNA提取试剂盒的使用方法步骤（离心机法）0、将Nuclease-Free Water 放在65℃的水浴锅中1、剪取动物组织20mg，分成相同的2份，放在1.5ml的离心管中，样品中决不能含有软骨组织，否则会阻塞柱子2、每份样品中添加275ul先前准备的消化液，确保消化液能够完全覆盖样品，如果不能覆盖，将样品再剪得小点3、把样品放到55℃的水浴锅或加热块中孵育16-18小时（过夜），蛋白酶K消化过夜后2000g离心，取上清液到1.5ml的离心管中。
4、向样品中添加250ul的Wizard SV Lysis Buffer,漩涡混匀5、处理后的产物尽快进行下面的操作，如果不能进行下面的操作，应该把它放在-70℃下保存，使用时应该冻融或者放在55℃温和解冻6、将收集管做上标记，将柱子放在收集管上，把整个1.5ml离心管内的溶解产物转移到柱子中。
13000g离心3分钟，目的是让基因组吸附在柱子中，如果仍有残留液体，可以重新13000g离心一分钟7、倒掉收集管中的液体，把柱子重新放回收集管上8、检查一下乙醇是否已经添加到Wizard SV Wash Solution中9、每个样品中添加650ul的Wizard SV Wash Solution（真空抽滤要加800ul），13000离心1分钟10、去除溶液，将柱子重新放置在集合管中11、重复9-10步3次，（一共需要洗4次）12、洗剂结束后，清空收集管中的水13000g离心2分钟13、去掉收集管，取相同数量的1.5ml的离心管做上标记，将柱子将在 1.5ml的离心管上，添加250ul65℃的Nuclease-FreeWater 到柱子中，室温孵育2分钟，13000g离心1分钟（要溶液，勿扔）14、再在柱子中添加250ul的Nuclease-Free water，室温孵育2分钟，13000g离心1分钟15、扔掉柱子，将收集的纯化的DNA储存在-20℃至-70℃。
全血细菌DNA试剂盒Cat. No. 5次制备300400550次制备3004050250次制备3004250核酸纯化柱2 ml离心管溶菌酶Buffer L1Buffer L2Buffer WA（浓缩液）Buffer WB（浓缩液）Buffer TE说明书5个5个30 mg2 ml2 ml1.9 ml1.5 ml1.2 ml1份50个50个300 mg16 ml16 ml12 ml10 ml12 ml1份250个250个1.5 g80 ml80 ml60 ml50 ml60 ml1份产品储存与有效期溶菌酶请置于2~8℃贮存。
其他物品如果储存于室温（15~25℃），可在两年内保持使用性能无明显变化；如果将产品储存于2~8℃，可延长产品的有效期至两年以上。
技术支持杭州新景生物产品介绍本产品适合从350 µl新鲜的或者是冷冻贮藏的人或动物全血中快速分离纯化血液总DNA。
溶菌酶将全血中的细菌破壁后，裂解液溶解全血及血液中的细菌，再经Buffer L2沉淀去除血红蛋白，上清中的细菌DNA与细胞DNA一起吸附到纯化柱上，经Buffer WA和Buffer WB洗涤去除残留在膜上的蛋白与PCR抑制物后，DNA用Buffer TE洗脱，并可立即用于各种分子生物学实验。
用户需自备的试剂与物品1．去离子纯水、无水乙醇2．1.5 ml离心管3．移液器吸头（为避免样品间的污染，建议选用含有滤芯的移液器吸头）4．一次性手套及防护用品和纸巾5．台式小量离心机（可配离心1.5 ml离心管和2 ml离心管的转子）6．水浴锅或干浴锅、旋涡震荡器使用前准备1.如果离心机有制冷功能，请将温度设置到25℃。
2.将水浴锅温度设置到37℃，并将Buffer TE温育至37℃。
3.根据一次提取的标本数（按每个标本需加50 µl溶菌酶溶液计算）配制适量的100 mg/ml的溶菌酶溶液：比如要提取10个标本的细菌基因组DNA，则称取55 mg溶菌酶干粉，加入550 µl去离子纯水配制成550 µl溶菌酶溶液。
DNase使⽤说明promegaProduct ContentsRQ1 RNase-Free DNase:Part No.Size (units)M610A1,000D e s c r i p t i o n:RQ1 (RNA-Qualified) RNase-Free DNase is a DNase I (endonuclease) that degrades both double-stranded and single-stranded DNA, producing 3′-OH oligonucleotides (1). (RQ1 RNase-Free DNase may be used in applications where maintaining the integrity of the RNA is critical.) This DNase is suited for applications such as nick translation (2), production of random fragments (3), cleavage of genomic DNA for footprinting (3), removal of DNA template after in vitro transcription (4), and removal of DNA from RNA samples prior to applications such as RT-PCR (5).In the presence of Mg2+, DNase I attacks each strand of DNA independently, and the sites of cleavage are distributed in a statistically random fashion (6). In the presence of Mn2+, DNase I cleaves both strands of DNA at approximately the same site to yield fragments with blunt ends or protruding termini of one or two nucleotides in length (6).10X Reaction Buffer (M198A): The RQ1 DNase 10X Reaction Buffer provided with this enzyme has a composition of400mM Tris-HCl (pH 8.0), 100mM MgSO4and 10mM CaCl2.Enzyme Storage Buffer: RQ1 DNase is supplied in 10mM HEPES (pH 7.5), 50% glycerol (v/v), 10mM CaCl2and10mM MgCl2.Heat Inactivation:10 minutes at 65°C in the presence of Stop Solution.Inhibitors:EGTA; EDTA (7); salt concentrations >100mM will reduce DNase activity.Molecular Weight: 31,000 Daltons.Requirement: Ca2+and Mg2+or Mn2+(7).Source: Bovine pancreas.Storage Temperature: Store at –20°C. Avoid exposure to frequent temperature changes. See the expiration date on the Product Information Label.Stop Solution (M199A): 20mM EGTA (pH 8.0).Unit Definition:One unit of RQ1 RNase-Free DNase is defined as the amount required to completely degrade 1µg of lambda DNA in 10 minutes at 37°C in 50µl of a buffer containing 40mM Tris-HCl (pH 7.9), 10mM NaCl, 6mM MgCl2and 10mMCaCl2. Under these assay conditions one unit of RQ1 DNase activity is approximately equal to one Kunitz unit. See the unit concentration on the Product Information Label.Usage Notes:1.This DNase solution does not contain an RNase inhibitor. Observe caution in handling the product to ensure against contaminating it with RNase.2.Under different buffer conditions the amount of DNase required to completely digest a given amount of DNA may need to beempirically determined. For example, salt concentrations >100mM will reduce DNase activity.Quality Control AssaysContaminant ActivityRNase Assay: To test for RNase activity,50ng of [3H]RNA is incubated with 5 units of RQ1 RNase-Free DNase in Transcription Optimized 1X Buffer (Cat.#P1181, diluted fivefold) for 1 hour at 37°C, and the release of radiolabeled nucleotides is monitored by scintillation counting of TCA-soluble material. The minimum passing specification is <3% release.Promega Corporation2800 Woods Hollow RoadMadison, WI 53711-5399USATelephone608-274-4330Toll Free800-356-9526Fax608-277-2516Internet/doc/372331592.htmlPRODUCT USE LIMITATIONS, WARRANTY,DISCLAIMER Promega manufactures products for a number ofintended uses. Please refer to the product label forthe intended use statements for specific products. Promega products contain chemicals which may be harmful if misused. Due care should be exercisedwith all Promega products to prevent direct human contact.Each Promega product is shipped with documentation stating specifications and other technical information. Promega products are warranted to meet or exceed the stated specifications. Promega's sole obligation and the customer's sole remedy is limited to replace-ment of products free of charge in the event products fail to perform as warranted. Promega makes no other warranty of any kind whatsoever, and SPECIFICALLY DISCLAIMS AND EXCLUDES ALL OTHER WAR-RANTIES OF ANY KIND OR NATURE WHATSOEVER, DIRECTLY OR INDIRECTLY, EXPRESS OR IMPLIED, INCLUDING, WITHOUT LIMITATION, AS TO THE SUITABILITY, PRODUCTIVITY, DURABILITY, FITNESS FOR A PARTICULAR PURPOSE OR USE, MER-CHANTABILITY, CONDITION, OR ANY OTHER MAT-TER WITH RESPECT TO PROMEGA PRODUCTS. Inno event shall Promega be liable for claims for any other damages, whether direct, incidental, foresee-able, consequential, or special (including but not lim-ited to loss of use, revenue or profit), whether based upon warranty, contract, tort (including negligence) or strict liability arising in connection with the sale or the failure of Promega products to perform in accordance with the stated specifications.Part# 9PIM610Revised 2/05Part# 9PIM610Printed in USA. Revised 2/05Riboprobe is a trademark of Promega Corporation and is registered with the U.S. Patent and Trademark Office.? 1996–2005 Promega Corporation. All Rights Reserved.All specifications are subject to change without prior notice.Product claims are subject to change. Please contact Promega Technical Services or access the Promega online catalog for the most up-to-date information on Promega products.AF9PI M6100205M610(a)The PCR process is covered by patents issued and applicable in certain countries. Promega does not encourage or support the unauthorized or unlicensed use of the PCR process. Use of this product is recommended for persons that either have a license to perform PCR or are not required to obtain a license.(b)U.S. Pat. No. 5,552,302, European Pat. No. 0 422 217, Australian Pat. No. 646803 and Japanese Pat. Nos. 3009458 and 3366596 have been issued to Promega Corporation for the methods and compositions for production of human recombinant placental ribonu-clease inhibitor.(c)U.S. Pat. Nos. 4,966,964, 5,019,556 and 5,266,687, Australian Pat. Nos. 616881 and 641261 and other pending and issued patents, which claim vectors encoding a portion of human placental ribonuclease inhibitor, are exclusively licensed to Promega Corporation.Usage Information。
