Combitips plus分液管,标准级,10 ml,100个

化学专业英语分析术语英语翻译玻璃漏斗 Glass funnel long stem试管 test tube test tube brush test tube holder test tube rack 蒸发皿 evaporating dish small烧杯 beaker锥形瓶 Erlenmeyer量筒 grad cylinder洗瓶 plastic wash bottle勺皿 casserole ,smallstoppered flask分液漏斗 separalory funnelwater bath/oil bathstrring barmagnetic stirrer冷凝器 condenserBallast bottle圆颈烧瓶 Round-buttom flask试剂瓶 reagent bottles托盘天平 platform balance 台秤0.1g 托盘pan 指针刻度表pointer and scalecrossbeams and sliding weights 游码分析天平 two-pan/single-pan analytical balance滴定管 burette glass bead(basic) nozzle移液管 pipette 胖肚 elongated glass bulb洗耳球 rubber suction bulb玻棒 glass rod玻璃活塞 stopcock容量瓶 pyknowmeter flasks比重瓶 (one-mark)volumetric flasks胖肚吸管 one-mark pipette刻度吸管 graduated pipettes实验仪器清单1、柜子中四、抽屉中：锥形瓶(conical flask) 250ml×4 药匙(medicine spoon)×1 (Erlenmeyer flask) 100 ml×3 滴管(drip tube;dropper)×2烧杯(beaker) 500 ml×1 玻棒(Glass stic)×2250 ml×3 木试管夹(test tube clamp;test tube holder)×1100 ml×3 胖肚吸管(straws) 25 ml×150 ml×2 10 ml×1容量瓶(volumetric flask) 100 ml×2 乳钵(morta)×150 ml×4 洗耳球(ear wadhing bulb)碘量瓶 (iodin numoe flask;iodineflask) 500 ml×3试剂瓶 (reagent bottle) （无色）×2（棕色）×2 配洗液：量筒（cylinder） 100 ml×1 K2Cr2O72g+5ml水→65mlH2SO4(graduated cylinder)10ml×1 边加边搅拌(stir)。

真空采血管的各种规格及应用之袁州冬雪创作血惯例管（Blood routine tube）血惯例管（Blood routine tube）用于临床血液学检查，交叉合血和血型鉴定等，并适合给类细胞分析仪，对血细胞全面周到的呵护，尤其对血小板的呵护作用，有效阻止血小板堆积，并呵护血细胞的形态和体积在较长的时间内不受影响，采取超维系平均的喷雾设备及工艺，使添加技能平均分散在管壁，与血液标本充分混匀.EDTA抗凝血浆用于病原微生物、寄生虫、细菌的分子生物学鉴定.促凝管（Pro-coagulation tube）用于医学检验中生化学、免疫学检查的血液标本的收集，它具有操纵温度范围广，内壁经特殊处理，极度光滑，高品质的促凝剂平均分散在采血管内壁上，血液标本进入管内后与其充分接触，在5—6分钟内使血液完全凝结，并通过离心获得高质量的血清，处理了血细胞的凝结破裂，溶血及纤维蛋白析出问题，知足疾速门诊、急诊血清生化试验.血块完全收缩时间为20分钟—25分钟，离心转速为3500—4000转/分，离心时间为5分钟，简易贮存温度4℃—25℃.分离胶管（Gel and clot activator tube）用于血清生化学、免疫学、药物检测等，内壁上平均涂布促凝剂，可大大缩短血液凝结时间由于引进的分离胶为纯净物，其物理化学性质较稳定，抗高温才能强，有效防止储运过程中，分离胶分解变形，离心后的分离胶固化形成屏障，将血清与血细胞完全分离，有效阻止二者间的物质交换，获得高质量的血清，使检测成果更加真实，提高血清的收集率，坚持血清稳定时间超出48小时，其生化性质与化学构成不发生显著变更，可直接上机.血块完全收缩时间为20分钟—25分钟，离心转速为3500—4000转/分，离心时间为5分钟，简易贮存温度4℃—25℃.肝素管（Hepar in tube）用于临床血浆、急诊生化、血液流变学测定的血液标本收集对血液成分干扰少，不影响红细胞体积，不引起溶血，具有血浆分离转速快，操纵温度范围广，与血清标本指标兼容性强，激活纤溶酶和抑制凝血活酶，使纤维蛋白原和纤维蛋白之间达成动态平衡，防止发生纤维蛋白丝血浆的大部分指标6小时内可重复检测.肝素锂除具有肝素钠的特性，还可以用于微量元素检测，而且对钠离子不发生影响.可按客户要求添加血浆分离胶，制备优良血浆血液知足临床试验需要.离心转速为3500—4000转/分，离心时间为3分钟，简易贮存温度4℃—25℃.无添加剂管（No additive tube）用于医学检验中生化学、免疫学、血清学、各种病毒监测，微量元素检测，血库检查血液标本的收集与盛装.内壁经特殊处理，极度光滑，使血小板活性正常，凝血顺畅，防止溶血及血细胞和纤维蛋白粘附在内壁的现象，为临床经历提供足量无污染血清标本，并能较长维持血清的正常成分及利于血清的复检，可重复性好.—2小时，离心转速为3500—4000转/分，离心时间为5分钟，简易贮存温度4℃—25℃.血凝管（PT tube）用于凝血项目标实验，适用于纤溶系统（凝血酶原时间，凝血酶时间，活化部分凝血酶时间，纤维蛋白等），按抗凝剂与血样的比例胃1:9设定，配比切确包管检验成果的有效性，防止误诊，由于枸橼酸钠毒性小，也用于血液保管.采血时务必采足血量，以包管检验成果的准确性.血沉管（ＥＳＲtube）专用于红细胞沉降速率的测定时学员可以的收集和抗凝.血沉管（ＥＳＲtube）适用于多种品牌全自动血沉仪配套使用.由于采血量少，关内负压较小，抽血时间相对较长，应耐烦等待直至血液停止流入采血管后混匀５—８次，使抗凝剂和血液充分混匀.不恰当的混匀会造成溶血＼凝血和血泡，影响检测效果.血糖管（Oxalate tube）用于血糖、耐糖量、红细胞电泳、抗碱血红蛋白、糖溶血和乳酸盐等项目检查的血样收集.由于抑制性抗凝剂的加入，有效阻止血糖代谢，处理了溶血的困难，使得血液的原始状态坚持时间长，确保了血糖24小时内检测数据恒定.添加剂有氟化钠+草酸钾、氟化钠+肝素钠、氟化钠+EDTAK2、氟化钠+EDTAN2.离心转速为3500—4000转/分，离心时间为3分钟，简易贮存温度4℃—25℃.条形码采血管（bar code tube）实现病人唯一的身份识别，包管了样本检验信息的准确性，使得样本在传递、检验、成果反馈的每个环节都可以通过条形码及相关信息确认病人身份.条形码的使用增加患者的信任度，提高医护人员的工作效率，同时降低了由于重复化验，诊疗所带来的费用，防止医患胶葛.有利于检验仪器操纵的规范化和功能操纵，提高实验室的自动化程度.。

Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Printed in USA.Part# TB173Revised 12/10Page 11.Description (1)2.Product Components and Storage Conditions (2)3.Production of a Cleared Lysate (2)4.Plasmid DNA Purification (4)5.Supplementary Information (6)A.Factors Affecting Plasmid DNA Yield (6)B.Choosing a Bacterial Strain (6)C.Special Considerations for Automated Fluorescent Sequencing (7)6.Troubleshooting (9)position of Buffers and Solutions (11)8.Related Products (12)9.References (12)1.DescriptionSmall-scale purifications of plasmid DNA, known as minipreps, are commonly used in molecular biology procedures. Over the years many miniprep protocols have been used, but few have proven to be consistently reliable (1). The strategy has been improved and adapted for processing larger culture volumes (10–100ml) with the Wizard ®Plus Midipreps DNA Purification System (a).The Wizard ®Plus Midipreps DNA Purification System eliminates many of the problems associated with standard midiprep procedures and provides a simple and reliable method for rapid isolation of plasmid DNA. This system can be used to isolate any plasmid but works most efficiently with plasmids <20,000bp.When using the standard protocol, the entire midiprep process can be completed in 90 minutes or less with no organic extractions or ethanol precipitations. Multiple midipreps can be easily processed at one time with the Vac-Man ®(20-sample capacity, Cat.# A7231) or Vac-Man ®Jr. (2-sample capacity, Cat.# A7660) Laboratory Vacuum Manifold. DNA is eluted from the Wizard Midicolumn in Nuclease-Free Water (Cat.# P1193). The purified plasmid can be used directly for automated fluorescent DNA sequencing or Wizard ®Plus Midipreps DNA Purification SystemAll technical literature is available on the Internet at /tbs Please visit the web site to verify that you are using the most current version of this Technical Bulletin. Please contact Promega Technical Services if you have questions on useof this system. E-mail techserv@.restriction digestion without further manipulation and also can be used forin vitro transcription reactions supplemented with a ribonuclease inhibitor, such as Recombinant RNasin®Ribonuclease Inhibitor (Cat.# N2511).2.Product Components and Storage ConditionsProduct Size Cat. # Wizard®Plus Midipreps DNA Purification System25 preps A7640 Each system contains sufficient reagents and columns for 25 isolations from 10–100ml of bacterial culture (using EndA– strains). Includes:•75ml Cell Resuspension Solution (CRA)•75ml Cell Lysis Solution (CLA)•150ml Neutralization Solution (NSA)•250ml Wizard®Midipreps DNA Purification Resin•355ml Column Wash Solution* (CWB)•25Midicolumns with Reservoirs*Column Wash Solution is provided in two bottles, one containing 125ml and the other 230ml of Column Wash Solution.Product Size Cat.# Wizard®Midipreps DNA Purification Resin*(a)1,000ml A7701 Wizard®Midicolumns100 each A7651Storage Conditions and Stability:Store the Wizard®Plus Midipreps DNAPurification System at room temperature (15–30°C). No refrigeration is required.Protect the resin from exposure to direct sunlight.3.Production of a Cleared LysateStart each Wizard®Plus Midiprep with a 10–100ml overnight culture of E. coli.DNA yields may vary between 10µg and 200µg depending on the volume ofbacterial culture, the plasmid copy number and the bacterial strain used. Up to200µg of high-copy-number plasmid DNA can be obtained from a 100mlculture. When isolating DNA from low-copy-number plasmids, it isrecommended to process 100ml of bacterial culture.Materials to Be Supplied by the User(Solution compositions are provided in Section 7.)•centrifuge capable of 10,000–14,000 × g•Miracloth™ (Calbiochem Corp. Cat.# 475855, filter paper (Whatman®#1, GFA or GFC) or an autoclaved coffee filter•ethanol (95%)Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Part# TB173Printed in USA. Page 2Revised 12/10Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Printed in USA.Part# TB173Revised 12/10Page 3Before you begin , dilute both bottles of Column Wash Solution (CWB) asfollows:Add 320ml of 95% ethanol to the large bottle (230ml) for a final volume of550ml; add 170ml of 95% ethanol to the small bottle (125ml) for a final volume of 295ml.Note: Throughout the remainder of this document Column Wash Solution(CWB), Cell Resuspension Solution (CRA), Cell Lysis Solution (CLA) andNeutralization Solution (NSA) are referred to simply as Column Wash Solution,Cell Resuspension Solution, Cell Lysis Solution and Neutralization Solution.1.Pellet 10–100ml of cells by centrifugation at 10,000 × g for 10 minutes at4°C. Pour off the supernatant and blot the tube upside down on a papertowel to remove excess liquid.pletely resuspend the cell pellet in 3ml of Cell Resuspension Solution.(To aid resuspension, manually disrupt the pellet with a 12-inch applicatorstick or by pipetting until no clumps are visible. Complete resuspension iscritical for optimal yields.)3.Add 3ml of Cell Lysis Solution and mix by inverting the tube four times.Do not vortex. The cell suspension should clear immediately.Note:Some bacterial cells are more resistant to lysis and may requireincubation for 3–5 minutes for efficient lysis. Additionally, culture volumes>50ml will take an extra 3–5 minutes to clear. The lysate may not appearcompletely clear, but do not extend lysis time beyond 3–5 minutes as thismay result in the formation of single-stranded DNA in the preparation.4.Add 3ml of Neutralization Solution and mix by inverting the tube 4 times.Alternatively , if using an EndA+ strain, add 6ml of NeutralizationSolution, mix by inverting the tube 4 times, and incubate the lysate at roomtemperature for 10 minutes. Proceed to Step 5.5.Centrifuge at 14,000 × g for 15 minutes at 4°C. If a tight pellet has notformed by the end of the centrifugation, centrifuge for another 15 minutes.6.Carefully decant the supernatant to a new centrifuge tube, avoiding thewhite precipitate. Alternatively, transfer the cleared supernatant by filteringit through Miracloth™ (Calbiochem Corp. Cat.# 475855), filter paper(Whatman ®#1, GFA or GFC) or an autoclaved coffee filter into the newcentrifuge tube. Proceed immediately to Section 4.4.Plasmid DNA PurificationMultiple Wizard ®Plus Midipreps can be easily processed simultaneouslywith the Vac-Man ®or Vac-Man ®Jr. Laboratory Vacuum Manifold, which is required for this procedure.Materials to Be Supplied by the User(Solution compositions are provided in Section 7.)•vacuum pump or vacuum aspirator capable of a vacuum of 15–18 inches of mercury (Hg)•vacuum manifold (i.e., Vac-Man ®(Cat.# A7231) or Vac-Man ®Jr. (Cat.# A7660) Laboratory Vacuum Manifold)•Nuclease-Free Water (Cat.# P1193) preheated to 65–70°C •optional:40% isopropanol/4.2M guanidine hydrochloridesolution (required for EndA+ strains—use only PromegaCat.# H5381)Thoroughly mix the Wizard ®Midipreps DNAPurification Resin before removing an aliquot.1.Add 10ml of resuspended Wizard ®Midipreps DNA Purification Resinto the DNA solution from Section 3. Swirl to mix.Note:Extended incubation of the resin and lysate is not necessary. Donot allow the resin to remain in contact with the lysate for longer than ittakes to load the Midicolumns.2.For each Midiprep, use one Midicolumn. Insert the Midicolumn tip intothe vacuum manifold port.3.Transfer the resin/DNA mixture into the Midicolumn. Apply a vacuumof at least 15 inches of Hg to pull the resin/DNA mix into theMidicolumn. When all of the sample has passed through the column,break the vacuum at the source.If using an EndA+ strain:a.The total volume of lysate and resin will exceed the column capacity by1ml. Therefore, all but 2–4ml of the resin/DNA should be applied to thecolumn. After the vacuum is applied and the column volume drops, theremainder of the lysate and resin can be added.b.Add 15ml of 40% isopropanol/4.2M guanidine hydrochloride solution(Section 7) to each column. Apply a vacuum continuously until 30seconds after all of the solution has flowed through the columns. Notethat this solution will flow through the column more slowly than thestandard Column Wash Solution. Proceed to Step 4.4.Add 15ml of Column Wash Solution to the Midicolumn and apply avacuum to draw the solution through the Midicolumn.Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Part# TB173Printed in USA.Page 4Revised 12/105.Break the vacuum at the source and add another 15ml of Column WashSolution to the Midicolumn. Reapply a vacuum to draw the solutionthrough the Midicolumn.Note:The Column Wash procedure may take up to 30 minutes.6.Dry the resin by continuing to draw a vacuum for 30 seconds after thesolution has been pulled through the column. Do not dry the resin formore than 30 seconds.Remove the Midicolumn from the vacuum source.Separate the Reservoir from the Midicolumn by breaking or cutting withsharp scissors as shown in Figure 1. Transfer the Midicolumn to a 1.5mlmicrocentrifuge tube. Centrifuge the Midicolumn at 10,000 × g in amicrocentrifuge for 2 minutes to remove any residual Column WashSolution. Transfer the Midicolumn to a new microcentrifuge tube.7.Add 300µl of preheated (65–70°C) Nuclease-Free Water to the Midicolumnand wait 1 minute. Elute the DNA by centrifuging the Midicolumn at10,000 × g8.9.eluted DNA.Figure 1. Location of the cut site for separating the Reservoir from the Midicolumn.Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Printed in USA.Part# TB173Revised 12/10Page 5Cut column here0991M A 08_3A5.Supplementary InformationPlasmid DNA can be purified from 10–100ml overnight cultures of E. coliwith the Wizard®Plus Midipreps System. The yield of plasmid will varydepending on a number of factors, including the volume of bacterial culture,plasmid copy number, type of culture medium and the bacterial strain. Theprotocol presented in this technical bulletin is for the isolation of plasmidDNA from E. coli.5.A.Factors Affecting Plasmid DNA YieldPlasmid copy number is one of the most important factors affecting yield ina given system. Copy number is determined primarily by the region ofDNA surrounding and including the origin of replication in the plasmid.This area, known as the replicon, controls replication of plasmid DNA bybacterial enzyme complexes. Some DNA sequences, when inserted into aparticular vector, can lower the copy number of the plasmid. In addition,excessively large DNA inserts can also reduce plasmid copy number.5.B.Choosing a Bacterial StrainEndonuclease I is a 12kDa periplasmic protein that degrades double-stranded DNA. This protein is encoded by the gene end A. The E. coligenotype end A1 refers to a mutation in the wildtype end A gene, whichproduces an inactive form of the nuclease. E. coli strains with this mutationin the end A gene are referred to as EndA negative (EndA–). Table 1 containsa list of EndA– and EndA+ E. coli strains. The wildtype is indicated asEndA+. Using the Wizard®Plus Midipreps System, high-quality DNA iseasily obtained from both EndA+ and EndA– strains. Special precautionsmust be taken when working with EndA+ strains to ensure the isolation ofhigh-quality DNA (2), including the use of several modified protocol steps,as indicated, and the use of a less rich growth medium (e.g., LB). Themodified protocol will eliminate most problems associated with thesestrains. However, the level of endonuclease I produced is strain-dependent,and the modified protocol may not totally exclude endonuclease I fromplasmid DNA prepared from very high endonuclease I-producing strains.Also note that the modified protocol requires the use of increased volumesof several of the supplied solutions and, as a result, you will be unable toperform as many isolations. In general, we recommend using EndA– strainswhenever possible.Figure 2 depicts DNA isolated from varying amounts of culture, using high-and low-copy-number plasmids and an EndA– E. coli strain with theWizard®Plus Midipreps DNA Purification System.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Part# TB173Printed in USA. Page 6Revised 12/10Figure 2. Plasmid DNA yield as a function of culture volume and plasmid copynumber, determined by use of the Wizard ®Plus Midipreps DNA PurificationSystem.Representative yields of a high-copy-number plasmid, pGEM ®-3Zf(+)Vector, and a low-copy-number plasmid, pBR322, prepared in E. coli strain DH5α™.Cultures were grown overnight at 37°C in 25ml increments of LB mediumcontaining 100µg/ml ampicillin.Table 1. List of EndA– and EndA+ Strains.5.C.Special Considerations for Automated Fluorescent SequencingFor the application of automated fluorescent sequencing, special considerationshould be given to the selection of plasmid type and E. coli strain to optimizeyield and plasmid quality. Optimal automated fluorescent sequencing resultsare routinely obtained by using high-copy-number plasmids and EndA–strains of E. coli .Note:For fluorescent sequencing applications, elute and store the DNA innuclease-free water.Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Printed in USA.Part# TB173Revised 12/10Page 725020015010050Culture Volume (ml)P l a s m i d Y i e l d (μg )1460M A 04_6APurified plasmid DNA must be within the proper concentration range forsuccessful automated cycle sequencing (ideally 0.2µg/µl, not less than0.1µg/µl).When working with plasmid DNA from low-copy-numberplasmids, we strongly recommend that DNA concentrations be determinedby agarose gel/ethidium bromide quantitation prior to any application.DNA quantitation by spectrophotometric methods is prone to errors andrequires a large amount of sample.The Wizard®Plus Midipreps System routinely yields 70µg of medium- orhigh-copy-number plasmid DNA when used with the pGEM®Vector andDH5α™ cells in 25ml of culture. For low-copy-number plasmids, a largerculture volume is required to obtain sufficient DNA for sequencing. Insome cases it may be possible to amplify plasmid DNA by growing thebacteria in the presence of antibiotics such as chloramphenicol orspectinomycin (1).Special Considerations for Sequencing Using BigDye®ChemistryWhen performing dilutions of BigDye™ terminator ready reaction mix, it isessential to use an appropriate dilution buffer, such as 250mM Tris-HCl (pH9.0), 10mM MgCl2.Table 2 outlines the amount of terminator-ready reaction mix and dilutionbuffer required to obtain the appropriate dilution for BigDye®terminatorreactions. For details on running these reactions, please refer to the protocolsupplied with the BigDye®terminator system. For each reaction, add thereagents in Table 2 to a separate tube.Table 2. Appropriate dilutions for BigDye™ Terminator Reactions.AmountComponent No Dilution1:21:41:6terminator-readyreaction mix*8.0µl 4.0µl 2.0µl1.3µldouble-strandedplasmid DNA template200–500ng200–500ng200–500ng200–500ngprimer 3.2pmol 3.2pmol 3.2pmol3.2pmoldilution buffer**0µl 2.0µl 3.0µl3.4µlNuclease-Free Waterto a final volume of20µl20µl20µl20µl**Dilution buffer is a 5X solution.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Part# TB173Printed in USA. Page 8Revised 12/106.TroubleshootingFor questions not addressed here, please contact your local Promega Branch Office or Distributor. Contact information available at: . E-mail: techserv@Symptoms Causes and CommentsPoor cell lysis Too many bacterial cells in culture medium.Use LB medium to grow bacteria. The use ofrich media or excessive culture volumes maylead to a biomass value too high for completelysis. All media should contain antibiotics at theappropriate concentration.Poor resuspension of bacterial cell pellet. Thecell pellet must be thoroughly resuspendedprior to cell lysis. Pipet or disperse (using anapplicator stick) the pellet with CellResuspension Solution. No cell clumps shouldbe visible after resuspension.No plasmid DNA purified Ethanol not added to the Column WashSolution. Prepare the Column Wash Solution asinstructed before beginning the procedure.EndA+ strain of bacteria used. DNA appearsdegraded or lost upon incubation with Mg2+containing buffer, i.e., restriction enzyme buffer.Follow protocol modifications for EndA+strains of bacteria.Inaccurate quantitation of plasmid DNA yield.Quantitate plasmid DNA yield by agarose gel/ethidium bromide electrophoresis.DNA floats out of well Carryover of residual ethanol from Columnduring loading of agarose gel Wash Solution. Follow directions forappropriate drying of resin by vacuum andcentrifugation. If DNA has already been eluted,precipitate DNA and dry remaining ethanolfrom the DNA pellet prior to resuspension inNuclease-Free Water. Increase loading dyeconcentration by 2X.Low plasmid DNA yields Overgrowth of bacterial culture by nontrans-formed bacteria. Make certain that antibioticswere used in all media, both liquid and solid.Do not culture bacteria longer than 24 hours.Optimal culture length is 12–16 hours.Bacterial culture too old. Inoculate antibiotic-containing media with freshly isolated bacterialcolony from an overnight plate.Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Printed in USA.Part# TB173 Revised 12/10Page 96.Troubleshooting (continued)Symptoms Causes and CommentsLow plasmid DNA yields (continued)Low-copy-number plasmid used. See Section5.A. Cultures should not exceed the maximumrecommended volumes per isolation.Precipitate formed in resin. Warm resin in 37°Cwater bath for 15–20 minutes. Gently swirl bottleto mix and allow to cool to 30°C prior to use.Resin fines in eluted DNA. Follow directions forremoval of resin fines from eluted DNA, i.e.,filtration and centrifugation. If DNA aggregatehas formed, heat in the presence of 1M NaCl toredissolve aggregate. Centrifuge to remove resinfines. Precipitate DNA with ethanol and washwith 70% ethanol to remove residual NaClbefore using in downstream applications.Overdrying of resin on vacuum source. Followdirections for drying on vacuum source. Do notdry for times longer than suggested.Wrong reagents used. Make certain that ColumnWash Solution is diluted with ethanol beforeuse. Note that Wizard®Plus and Wizard®PlusSV components are not interchangeable.Plasmid DNA yield not accurately quantitated.Use agarose gel/ethidium bromide quantitation.Nicking of plasmid DNA Overincubation during the alkaline lysis step.Total incubation of cell suspension with CellLysis Solution should not exceed 5 minutes.No results or poor results with Too little DNA was added to the sequencingautomated fluorescent sequencing reaction. Inoculate fresh LB medium with anewly isolated E. coli colony. Purify plasmidDNA and quantitate by agarose gel/ethidiumbromide electrophoresis.TE buffer was used for DNA elution. Ethanolprecipitate and resuspend pellet in Nuclease-FreeWater. (The EDTA in TE buffer can interferewith downstream applications by chelating Mg2+.)ABI PRISM®BigDye®chemistry was used. Use ofthis chemistry necessitates ethanol precipitationof eluted DNA prior to sequencing reaction.Plasmid concentration not accurately quantitated.Use agarose gel/ethidium bromide electro-phoresis to accurately quantitate plasmid DNA. Promega Corporation·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 · Part# TB173Printed in USA. Page 10Revised 12/106.Troubleshooting (continued)SymptomsCauses and CommentsNo restriction digestionIncrease the amount of restriction enzyme used and/or the length of incubation time. Digest at the recommended temperature and in the optimal buffer for the restriction enzyme used.DNA degraded during restriction digestion due to use of EndA+ E. coli strain. Repurify DNA from fresh culture containing antibiotics. Follow instructions (Section 3 and 4) for EndA+ strains or use an EndA– strain of E. coli .Genomic DNA contaminationVortexing or overmixing after addition of the Cell Lysis Solution. Do not vortex samples after addition of Cell Lysis Solution to prevent shearing of genomic DNA.DNA yields on gel look low compared Traces of contaminants may be present in the to spectrophotometer readingseluted DNA, inflating the spectrophotometer readings. Phenol:chloroform extract andprecipitate DNA, then wash with 70% ethanol before repeating spectrophotometer readings. Alternatively, quantitate DNA by agarose gel/ ethidium bromide electrophoresis.7.Composition of Buffers and SolutionsPromega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·Printed in USA.Part# TB173Revised 12/10Page 11Cell Resuspension Solution (CRA)50mM Tris-HCl (pH 7.5)10mM EDTA 100µg/ml RNase A Cell Lysis Solution (CLA)0.2M NaOH 1%SDSNeutralization Solution (NSA)1.32M potassium acetate (pH 4.8)Column Wash Solution (CWB)80mM potassium acetate 8.3mM Tris-HCl (pH 7.5)40µM EDTA Add 320ml and 170ml of 95% ethanol to the large and small bottles,respectively, of Column Wash Solution (Section 3). Final ethanol concentration will be approximately 55%.(Component concentrations listed are for final solution with ethanol added.)TE buffer (1X)10mM Tris-HCl (pH 7.5)1mM EDTA 40% isopropanol/4.2M guanidine HCl66.9g guanidine hydrochloride (use only Promega Cat.# H5381)Prepare a 7M solution by dissolving the guanidine hydrochloride in 50–60ml of sterile, distilled water. This reaction is very endothermic; warming the mixture to 37°C (do not exceed 37°C) will speed the process. Bring to a final volume of 100ml with sterile, distilled water.Prepare the 40% isopropanol/4.2M guanidine HCl solution by combining 30ml of the 7M guanidine HCl solution with 20ml of isopropanol in a 50ml screw-cap tube and mixing thoroughly.Store at room temperature.(a)U.S. Pat. Nos. 5,658,548 and 5,808,041, Australian Pat. No. 689815 and European Pat. No. 0 723 549 have been issued toPromega Corporation for nucleic acid purification on silica gel and glass mixtures. Other patents are pending. © 2010 Promega Corporation. All Rights Reserved.pGEM, RNasin, Vac-Man and Wizard are registered trademarks of Promega Corporation. PureYield is a trademark of Promega Corporation.ABI PRISM and BigDye are registered trademarks of Applera Corporation. Luer-Lok is a registered trademark of Becton-Dickinson & Co. DH5αis a trademark of Life Technologies, Inc. Miracloth is a trademark of Calbiochem Corporation. Whatman is a registered trademark of Whatman Paper Company, Ltd.Products may be covered by pending or issued patents or may have certain limitations. Please visit our Web site for more information.All prices and 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.8.Related ProductsProductSize Cat.#PureYield™ Plasmid Midiprep System25 preps A2492100 preps A2495Wizard ®Plus SV Minipreps DNA Purification System 50 preps A1330(does not include Vacuum Adapters)250 preps A1460Cell Resuspension Solution 150ml A7112Cell Lysis Solution 150ml A7122Neutralization Solution 150ml A7131Column Wash Solution125mlA8102ProductSizeCat.#Vac-Man ®Laboratory Vacuum Manifold 20-sample capacity A7231Vac-Man ®Jr. Laboratory Vacuum Manifold 2-sample capacityA7660One-Way Luer-Lok ®Stopcocks10 eachA72619.References1.Ausubel, F.M. et al.(1989) Current Protocols in Molecular Biology , Vol. 2, John Wiley & Sons, New York.2.Schoenfeld, T. et al.(1995) DNA purification: Effects of bacterial strains carrying the end A1 genotype on DNA quality isolated with Wizard ®plasmid purification systems. Promega Notes 53, 12.Promega Corporation ·2800 Woods Hollow Road ·Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526·Phone 608-274-4330 ·Fax 608-277-2516 ·Part# TB173Printed in USA.Page 12Revised 12/10。

试管test tube试管架test tube holder漏斗funnel分液漏斗separatory funnel烧瓶flask锥形瓶conical flask烧杯beaker天平balance/scale分析天平analytical balance酒精灯alcohol burner量筒graduated flask/measuring cylinder 洗瓶plastic wash bottle滴定管burette冷凝器condenser试剂瓶reagent bottles台秤platform balance容量瓶volumetric flask/measuring flask 移液管(one-mark) pipette刻度移液管graduated pipettes洗耳球rubber suction bulb玻棒glass rod蒸馏烧瓶distilling flask坩埚crucible表面皿watch glass称量瓶weighing bottle研磨钵mortar研磨棒pestle滴瓶dropping bottle小滴管dropper蒸馏装置distilling apparatus蒸发器evaporator铁架台iron support移液管架pipet rack试管架tube rack沸石boiling stone橡胶管rubber tubing药匙lab spoon镊子forceps坩埚钳crucible tong剪刀scissor打孔器stopper borer石棉网asbestos-free wire gauze电炉丝wire coil for heater脱脂棉absorbent cottonphph试纸universal ph indicator paper滤纸filter paper称量纸weighing paper擦镜纸wiper for lens秒表stopwatch量杯glass graduates with scale水银温度计mercury-filled thermometer ph计ph meter真空泵vacuum pump冷、热浴bath离心机centrifuge口罩respirator磁力搅拌器magnetic stirrer电动搅拌器power basic stirrer烘箱oven闪点仪flash point tester电炉heater。

SampleHandling• Mixers• Temperature control systems• Plates and reaction tubes• Centrifuges• Thermocyclers• PCR plates and tubes• 核酸蛋白测定仪• 比色皿I n s t r u m e n t s | S a m p l e H a n d l i n g产品应用产品特性其他不同电压制式的货号，请参见附录第 320 页。

更多相关信息请查询：/biophotometerplus定量更轻松：更多分光定量信息，详见第 287 页。

Technical specifications subject to change.‡多款预设测定方法用于核酸和蛋白定量以及荧光染料 标记率检测等‡340 nm ，405 nm 和 490 nm 可进行自定义检测，实现 高灵活性‡自动计算样品稀释度‡单波长检测，不涉及任何计算‡储存最近的 100 个结果及所有相关数据 ‡简单用户操作指南，避免误操作 ‡设计小巧，坚固耐用 ‡开机即可使用，无需预热 ‡测定时间短，操作简便‡无需外联电脑，单机即可操作 ‡氙灯光源，使用寿命长，光强度高的氙灯光源，仅在检测时段开启发射， 开机即用，无需预热，是一款节能高效 可循环材料 300 页。

BioPhotometer plus 核酸蛋白测定仪是专为分子生物学，生物化学和细胞生物学实验室设计的精巧型紫外/可见光分光光度计。

它提供32种常规快速检测方法，其中有9种可以自由编程。

只需按下一个按键，即可进行检测和显示计算结果。

检测结果和所有数据均将全屏显示，一目了然，保证安全、无差错操作。

BioPhotometer plus 外型精致轻巧，金属外壳坚固耐用，且便于携带和清洁。

不但可以适配比色皿使用，如Eppendorf 公司的UVette ®塑料比色皿，也可适配微量比色皿进行微量样品检测。

说明书Pierce™交联磁珠式免疫沉淀/免疫共沉淀试剂盒888052309.4货号描述88805 Pierce交联磁珠式免疫沉淀/免疫共沉淀试剂盒，包含足够完成40个反应的试剂，每个反应使用25 µL磁珠试剂盒组成:Pierce 蛋白 A/G 磁珠1 mL，贮存于含有 0.05% NaN3的水中，浓度为10 mg/mLPierce 免疫沉淀裂解/漂洗缓冲液，2 × 50 mL，pH 7.4，0.025M Tris，0.15M NaCl，0.001M EDTA，1% NP40，5% 甘油20×交联缓冲液，25 mL, 稀释后含10 mM 磷酸钠， 150 mM NaCl；pH 7.2DSS（辛二酸二琥珀酰亚胺酯），No-Weigh™形式， 8 × 2 mg 微型管中和液，1.0 mL，pH 8.5洗脱液，25 mL，pH 2.0电泳上样缓冲液，非还原性，（5×）, 5 mL，pH 6.8，0.3M Tris·HCl，5% SDS，50%甘油，泳道标记示踪染料贮存条件：收到本产品后于4°C贮存。

冰盒运输。

目录产品简介 (2)流程简述 (2)重要产品信息 (2)所需额外试剂和仪器 (3)Pierce经典磁珠式免疫沉淀试剂盒流程 (3)A. 将抗体结合于蛋白A/G磁珠上 (3)B. 交联结合的抗体 (4)C．哺乳动物细胞裂解 (4)D. 手动抗原免疫沉淀 (5)E. 自动免疫沉淀 (5)问题解决 (7)网站附加信息 (7)Thermo Scientific KingFisher 仪器的常见问题 (8)Thermo Scientific相关产品 (8)产品简介Thermo Scientific™ Pierce™ 交联磁珠式免疫沉淀/免疫共沉淀试剂盒通过共价键将抗体交联于Thermo Scientific™ Pierce™ 蛋白 A/G磁珠上能够高效完成抗原免疫沉淀（IP）及免疫共沉淀（Co-IP）实验。

分离后水平离心分离前血浆层细胞层分离液层红细胞层FicollPlus 1.083货号：P4360规格：200mL产品简介：本产品是一种无菌、低内毒素水平的密度梯度分离液。

其分离原理是根据血细胞的密度差异，通过离心使细胞按相应密度梯度分布，从而将细胞从血液中分离出来。

产品指标：密度1.083±0.001g/mL 渗透压290~350mOsm/kg H2O 无菌0.1μm 滤膜过滤保存条件：本产品对光敏感，应该室温避光储存，保质期2年。

无菌开封后，保存于室温。

操作步骤：1.取新鲜抗凝全血（EDTA、枸橼酸钠或肝素抗凝均可）或者去纤维蛋白血液，用等体积的全血及组织稀释液或者PBS稀释全血。

2.在离心管中加入适量分离液（当稀释后血液体积小于3mL时，加入3mL分离液；大于等于3mL，加入等体积分离液。

但二者的总体积不能超过离心管的三分之二，否则会影响分离效果），将稀释后的血液平铺到分离液液面上方，注意保持两液面界面清晰。

（可以使用巴氏德吸管吸取血液，然后将血液小心的平铺于分离液上，因为两者的密度差异，将形成明显的分层界面。

如果样品较多加样时间较长，在离心之前出现红细胞成团下沉属正常现象。

）3.室温，水平转子500~1000g，离心20~30min（血液的体积越大所需的离心力越大，离心时间越长，最佳的分离条件需摸索，离心转速最大不超过1200g）。

4.离心后将出现明显的分层：最上层是稀释的血浆层，中间是透明的分离液层，血浆与分离液之间的白膜层即为细胞层，离心管底部是红细胞与粒细胞。

5.小心的吸取白膜层细胞到15mL洁净的离心管中，10mL PBS或细胞洗涤液洗涤白膜层细胞。

250g，离心10min。

6.弃上清，5mL的PBS或细胞清洗液重悬细胞，250g，离心10min。

7.重复步骤68.弃上清，细胞重悬备用。

注意事项：A.开封前颠倒混匀，本分离液为无菌产品，为延长分离液保存时间，请在无菌条件下启封，避免微生物污染。

本生新品推荐：分子生物试剂-转膜—缓冲液系列
新品推荐：分子生物试剂-转膜—缓冲液系列
缓冲液：缓冲溶液指的是由弱酸及其盐、弱碱及其盐组成的混合溶液，能在一定程度上抵消、减轻外加强酸或强碱对溶液酸碱度的影响，从而保持溶液的pH值相对稳定。

分子生物试剂-转膜—缓冲液系列本生一直视质量控制为企业的生命，追求企业竞争力的不断提升。

公司在经营中始终秉承：遵纪守法，严于律己，宽仁以待，敢于承担的企业精神作为标准，以过硬的质量和优良的服务来维护和拓展市场，较大限度的满足客户的需求。

与客户的共赢，是我们的发展目标。

本生!您信任的合作伙伴。

我们愿与您真诚合作，共创美好的未来。