Highlights
- Quick & Easy: Simple 20 minute procedure.
- Highest Yield: Recover 3x more DNA.
- Ultra-Pure: Ready for qPCR, Next-Gen Sequencing, arrays, etc
Description
| Elution Volume | ≤ 10 µl |
|---|---|
| Equipment | Water bath or heat block (55°C), microcentrifuge, and vortex. |
| Purity | High quality DNA is ready for all sensitive downstream applications such as PCR, endonuclease digestion, Southern blotting, genotyping, Next-Gen sequencing, bisulfite conversion, etc. (A260/A230≥ 2.0). |
| Size Range | Capable of recovering genomic and mitochondrial DNA sized fragments > 50 kb. If present, parasitic, microbial, and viral DNA will also be recovered. |
| Workflow | Utilize a Proteinase K Digestion and Zymo-Spin Column for effective recovery of DNA. |
| Yield | The DNA binding capacity of the well is 5 µg. Typically, mammalian tissues yield: 1-3 µg DNA per mg skeletal, heart, lung, and brain tissues and 3-5 µg DNA per mg liver and kidney. Human whole blood will yield 3-7 µg DNA per 100 µl blood sampled. |
Q1: What is the difference between the digestion buffers? Can one buffer be used for all sample types?
The BioFluid & Cell Buffer (Red) is designed to allow for rapid Proteinase K digestion with easy-to-lyse samples (e.g. mammalian cells and biological fluids).The Solid Tissue buffer (Blue) can be used for any sample type and requires a longer PK digestion time compared to using the BioFluid & Cell Buffer (Red).
Q2: Can Quick-DNA process crude lysates?
Yes, add 4 volumes of Genomic Lysis Buffer to 1 volume of crude lysate, homogenized, or digested sample (see Cell Suspensions and Proteinase K Digested Samples) and proceed with the remainder of the protocol.
Q3: What is the purpose of adding beta-mercaptoethanol? Can this step be substituted or omitted?
Beta-mercaptoethanol is a reducing agent that helps break down proteins and improves DNA recovery and purity. Addition of beta-mercaptoethanol is recommended to enhance sample lysis, but can be substituted with dithiothreitol (DTT, final concentration of 10 mM) or omitted.
Q4: Can I use Quick-DNA Plus to clean-up previously isolated DNA?
No, the kit is designed for direct use with biological samples. For clean-up of previously isolated DNA, please use the Genomic DNA Clean & Concentrator or the DNA Clean & Concentrator kits.
Q5: What are the expected yields for each sample type?
Keep in mind that there is sample to sample variability.
| Sample Type | Input Amount | Expected Yield |
|---|---|---|
| Eukaryotic Cells | 1x106 Cells | 5-6 µg |
| Skeletal, Heart, Lung, Brain Tissue | 1 mg | 1-3 µg |
| Liver and Kidney Tissue | 1 mg | 3-5 µg |
| Human Whole Blood | 100 µl | 5-7 µg |
Q6: I’m seeing some yield inconsistencies with my blood samples, what’s happening?
White blood cells, which are the major source of genomic DNA in blood, easily and quickly settles. Mix the blood sample well prior to aliquoting for purification.
Q7: Can Proteinase K digestion be performed overnight in DNA/RNA Shield?
Yes, samples can be digested overnight. Make sure to follow the appendix (page 8 appendix B) for processing liquids samples in DNA/RNA Shield and incubate at room temperature.
Q8: What is the difference between Quick-DNA and Quick-DNA Plus kits?
The Quick-DNA is optimized for cells, soft tissues, and homogenized/digested samples using a single lysis/binding buffer. The Quick-DNA Plus kits contain an optimized Proteinase K for processing a wider variety of sample inputs, such as cells, blood, tissues, etc. The upgraded Quick-DNA Plus recovers more DNA with higher purity compared to the Quick-DNA Kits.
Q9: Can the Quick-DNA Plus kit be used with bacterial samples?
E.coli cells are easy-to-lyse and can be processed directly with the Biological Fluids & Cells protocol. For an all-inclusive kit with any type of microbes (including tough-to-lyse), use any of Zymo Research’s Environmental Kits (e.g. Quick-DNA Fungal/Bacterial, Quick-DNA Fecal/Soil, ZymoBIOMICS DNA, etc.).
| Cat # | Name | Size | Price | |
|---|---|---|---|---|
| D3004-2-50 | g-DNA Wash Buffer | 50 ml | $18.00 | |
| D3004-4-10 | DNA Elution Buffer | 10 ml | $14.00 | |
| D3004-5-15 | DNA Pre-Wash Buffer | 15 ml | $10.00 | |
| D4068-3-25 | Genomic Binding Buffer | 25 ml | $28.00 | |
| D4068-2-6 | Solid Tissue Buffer (Blue) | 6 ml | $12.00 | |
| C1001-50 | Collection Tubes | 50 Pack | $15.00 | |
| C1103-50 | Zymo-Spin IC-XM Columns | 50 Pack | $63.00 |
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GenomeWalkerTM Universal Kit价格多少,哪里有卖的?
2.腹水浓稀释于3倍体积Tris缓冲液1~5 ml/min 速度于蛋白A-Sepharose柱用Tris缓冲液洗柱直至所未结合蛋白都洗脱采用A280监测
3.依用2~3倍柱床体积柠檬酸缓冲液、乙酸缓冲液液及甘氨酸·Cl缓冲液连续洗脱结合蛋白洗脱蛋白直接接入装缓冲液管缓冲液体积应所收集蛋白体积1/4
4.采用ELISA检测抗原特异性MAb
5.超滤器浓缩所离单克隆抗体1~5mg/ml所用超滤膜XM50单克隆抗体存于-20℃
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建议:做肝功能检查再结合肝功能情况遵医嘱进行抗病毒治疗
盐沉淀
Epicentre(Illumina旗公司)MasterPure™ Complete DNA Purification Kit采用种专利盐沉淀系列试剂盒覆盖各种品类型包括血液、固定组织、植物、酵母、昆虫等短短30钟内带高产量核酸损失极少
据Epicentre资深产品经理Cristine Kinross介绍些试剂盒用户测序或其物应用品制备前需再进行额外纯化除蛋白及其细胞碎片引入些必须除毒性化合物避免洗涤并改善核酸产量
Kinross谈道:利用MasterPure收核酸色纯度让品直接进入测序文库制备于其应用包括芯片、qPCR克隆适合外试剂盒能用于RNA纯化获总核酸经DNase处理能总RNA
种试剂盒
Promega提供广泛核酸纯化试剂盒采用几种同策略例Wizard® Genomic DNA Purification Kit基于经典沉淀全血、组织培养细胞、物及植物组织、酵母细菌提取基组DNA灵高效;ReliaPrep™ 系列试剂盒采用柱提简单快速外Promega提供基于磁珠纯化试剂盒
Promega公司基组全球产品经理Eric Vincent表示:尽管溶液确切组同裂解、结合、洗涤洗脱基本程适用于同结合(硅胶或纤维素)同纯化形式(纯化柱或磁珠)及手自纯化操作于些非重要品类型(FFPE品)需要些专门操作或处理
我发独特结合/洗涤缓冲液能效除抑制游反应物质优化系统让研究员能够捕获少量核酸并体积(30 μl )洗脱我发自化解决案适合各种试剂通量Vincent说
些试剂盒提供完整解决案能够极体积洗脱核酸同避免抑制剂残留洗脱液经些试剂盒纯化核酸用于PCR、Sanger测序、新代测序、芯片等游析
经典硅胶膜
QIAGEN核酸纯化试剂盒依赖DNA与纯化柱硅胶膜结合抑制剂洗掉完整DNA随柱洗脱产量高纯度高带更高质量新代测序文库
Marco Polidori - QIAGEN本制备全球产品经理 - 认处理极少量品或困难品FFPE高产量变格外关键FFPE品所产DNA能现随机胞嘧啶脱氨事件导致单核苷酸态性(SNP)QIAGENGeneRead DNA FFPE Kit能够除脱氨胞嘧啶避免NGS实验产错误结产品尚未式推若兴趣试用填写资料
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离液盐
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据介绍试剂盒综合硅胶膜结合微量离形式处避免昂贵树脂、乙醇沉淀及害机物苯酚、氯仿更重要纯化所DNA应用于限制性内切酶消化、PCR、Southern blot、测序等
(作者:James Netterwald/物通编译)
第十二条物制品产环境空气洁净度级别应与产品产操作相适应厂房与设施应原料、间体品造污染
第十三条产程涉及高危操作其空气净化系统等设施应符合特殊要求
第十四条物制品产操作应符合表规定相应级别洁净区内进行未列操作参照表适级别洁净区内进行:
洁净度级别
物制品产操作示例
B级背景局部A级
附录菌药品非终灭菌产品规定各工序
灌装前经除菌滤制品其配制、合并等
C级
体外免疫诊断试剂阳性血清装、抗原与抗体装
D级
原料血浆合并、组离、装前巴氏消毒
口服制剂其发酵培养密闭系统环境(暴露部需菌操作)
酶联免疫吸附试剂等体外免疫试剂配液、装、干燥、内包装
第十五条产程使用某些特定物体阶段应根据产品特性设备情况采取相应预防交叉污染措施使用专用厂房设备、阶段性产式、使用密闭系统等
第十六条灭疫苗(包括基重组疫苗)、类毒素细菌提取物等产品灭交替使用同灌装间灌装、冻干设施每装应采取充污染措施必要应进行灭菌清洗
第十七条卡介苗结核菌素产厂房必须与其制品产厂房严格产涉及物产设备应专用
第十八条致病性芽胞菌操作直至灭程完前应使用专用设施炭疽杆菌、肉毒梭状芽胞杆菌破伤风梭状芽胞杆菌制品须相应专用设施内产
第十九条其种类芽孢菌产品某设施或套设施期轮换产芽胞菌制品任何间能产种产品
第二十条使用密闭系统进行物发酵同区域同产单克隆抗体重组DNA制品
第二十条菌制剂产加工区域应符合洁净度级别要求并保持相压;操作致病作用微物应专门区域内进行并保持相负压;采用菌工艺处理病原体负压区或物安全柜其周围环境应相压洁净区
第二十二条菌(毒)操作区应独立空气净化系统自病原体操作区空气循环使用;自危险度二类病原体操作区空气应通除菌滤器排放滤器性能应定期检查
第二十三条用于加工处理物体产操作区设备应便于清洁污染清洁污染效性应经验证
第二十四条用于物体培养设备应能够防止培养物受外源污染
第二十五条管道系统、阀门呼吸滤器应便于清洁灭菌宜采用线清洁、线灭菌系统密闭容器(发酵罐)阀门应能用蒸汽灭菌呼吸滤器应疏水性材质且使用效期应经验证
第二十六条应定期确认涉及菌毒种或产品直接暴露隔离、封闭系统泄漏风险
第二十七条产程病原体污染物品设备应与未使用灭菌物品设备并明显标志
第二十八条产程需要称量某些添加剂或(缓冲液)产区域存放少量物料
第二十九条洁净区内设置冷库恒温室应采取效隔离防止污染措施避免产区造污染
面GMP附录3要求关键看体外试剂哪类般没问题
1.相同房间产段产
2.同房间产且使用同空调系统

