计算溶液所需的质量、体积或浓度。
为什么选择此级别
，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
室温。常规运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
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产品简介
该试剂盒提供了一种简单、快速、高效的提取DNA方法，适用于土壤和粪便样本。 在高盐存在时， DNA结合于硅基包被的Magbeads表面。漂洗后，高纯度的DNA被洗 脱于Buffer EBL或去离子水中。纯化得到的DNA纯度好（A260/280的比值在1.7-1.9之间）， 完整度高（>15 kb），可用于二代测序、定量PCR、芯片检测等下游实验。
该试剂盒可与CWE2100或CWE3200 32通道核酸提取仪和CWE9600或CWE960 96通道核酸提取仪进行匹配使用，简单、快速地进行高通量提取，大大降低了实验者 的工作量和实验中的人为误差。
自备仪器、试剂
1. 恒温混匀仪
2. 2/15 mL磁力架
3. 32通道核酸提取仪
4. 96通道核酸提取仪
5. 96 DW Plate
6. 8 channel Comb
7. Spin tips pack
8. 无水乙醇，异丙醇
9. 涡旋振荡仪或组织研磨仪
注意事项
1. 第一次使用前按照试剂瓶标签向Buffer GW1和Buffer GW2中加入无水乙醇并做好标 记。
2. Magbeads严禁冰冻、离心。冰冻、离心可能会对Magbeads造成不可逆的损害。
操作步骤
1. 样本前处理
1.1 短暂离心Lysis Tube以使珠子沉淀在底部。
1.2 A. 向 Lysis Tube中加入0.1-0.3g土壤或粪便样本，加入740-820 μL Buffer QSL与4 μLRNase A，旋紧管盖，短暂涡旋以混合。
B.若为非裂解型粪便保存液保存的粪便样本，向Ly- sis Tube中加入200 μL- 600 μL固液混合物，13000 rpm离心1 min，弃掉保存液 （若离心后的固体量过少，可再次富集，但不宜超过 0.3 g ）。加入620 μL Buffer QSL和4 μL RNase A，旋紧管盖，短暂涡旋以混合。
1.3 将Lysis Tube固定在装有2mL适配器的振荡研磨装置中，并根据您的设备使用优 化的研磨条件进行处理（参见附录）。
1.4 将Lysis Tube在恒温混匀仪上以70℃,1200rpm振荡10 min。随后13000 rpm离 心2 min以沉淀固体颗粒。转移540 μL上清液至新的2 mL离心管。
1.5 加入180 μL Buffer RIL，涡旋5 sec ，13000 rpm离心2 min。 注意： Buffer RIL待即将使用前取出，用完立即放在 2-8℃储存。
1.6 手动提取按第2部分操作，全自动核酸提取仪按第3或4部分操作。
2. 手动提取
2.1 在离心管中依次加入160 μL Buffer ML、480 μL 步骤 1.5 的上清液、320 μL 异丙醇、 20 μL Magbeads SN，涡旋振荡混匀5 sec后将离心管放于25℃、 1600 rpm的恒温 混匀仪上振荡混匀10 min。
注意： Magbeads SN加入前需涡旋振荡 20秒使其充分混匀。 Magbeads SN可与异丙醇按照上述 体积根据样本数量预混然后加入。
2.2 将离心管放于磁力架上静置1 min，待Magbeads完全吸附于离心管侧壁后彻底弃 去溶液（保持离心管固定于磁力架上）。
2.3 将离心管从磁力架上取下，加入750 μL Buffer GW1（使用前请检查是否已加入无 水乙醇）后涡旋点震1 min或涡旋振荡5 sec后放于25℃、 1600 rpm的恒温混匀仪 上振荡混匀2 min（振荡过程中确保Magbeads处于混匀状态）。之后将离心管放 于磁力架上静置1 min，待Magbeads完全吸附于离心管侧壁后轻轻颠倒磁力架将 离心管盖上的杂质洗落后彻底弃去溶液（保持离心管固定于磁力架上）。
2.4 重复步骤2.3。
2.5 将离心管从磁力架上取下，加入750 μL Buffer GW2（使用前请检查是否已加入无 水乙醇）后涡旋点震1 min或涡旋振荡5秒钟后放于25℃、 1600 rpm的恒温混匀仪 上振荡混匀2 min（振荡过程中确保Magbeads处于混匀状态）。之后将离心管放 于磁力架上静置1 min，待Magbeads完全吸附于离心管侧壁后轻轻颠倒磁力架将 离心管盖上的杂质洗落后彻底弃去溶液（保持离心管固定于磁力架上）。
2.6 重复步骤2.5。
2.7 保持离心管固定于磁力架上，用移液器进一步去除离心管管底和管盖上的溶液， 之后室温放置5-10分钟，使乙醇挥发干净（肉眼观察磁珠表面变成哑光且磁珠无 干裂）。
2.8 将离心管从磁力架上取下，加入50 -200 μL Buffer EBL。涡旋振荡使磁珠完全悬 浮于洗脱液中后将其放于56℃、 1600 rpm的恒温混匀仪上振荡洗脱10 min，或将 离心管放于56℃水浴锅中孵育10 min，期间每隔3 min涡旋振荡10 sec。
2.9 将离心管放于磁力架上静置2 min，待Magbeads完全吸附于离心管侧壁后用移液 器将洗脱液转移至新的离心管中-20℃保存备用。
3. 与32通道核酸提取仪匹配
3.1 按下表向96DW深孔板中加入相应试剂。
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注意：
1. 1&7 Colume中试剂按照顺序依次加入。
2. GW1、GW2使用前请检查是否已加入无水乙醇。
3. Buffer GW2与Magbeads SN 可按照上述体积根据样本数量预混，使用前涡旋 10 sec混匀。
3.2 将加入试剂的深孔板和磁套放于32通道核酸提取仪的相应位置，运行提取程序。约40 min后程序运行结束。
3.3 将深孔板6&12列中的洗脱产物转移至1.5 mL离心管中-20℃保存。
4. 与96通道核酸提取仪匹配
4.1 按下表向96DW深孔板中加入相应试剂。
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注意：
1. Plate1中试剂按照顺序依次加入。
2. GW1 、GW2使用前请检查是否已加入无水乙醇。
3. Buffer GW2与Magbeads SN可按照上述体积根据样本数量预混，使用前涡旋 10 sec混匀。
4.2 将加入试剂的深孔板和磁套放于CWE960（磁套置于盘位4）或CWE9600（磁套 置于盘位8）的相应位置，运行提取程序，约50 min后程序运行结束。
4.3 将深孔板Plate 6中的洗脱产物转移至1.5 mL离心管中-20℃保存。
附录：
使用以下方法之一研磨样本：
1. 在涡旋振荡仪上以最大速度手动涡旋振荡10 min。
2. 在搭配有1.5-2 mL水平离心管支架的涡旋振荡仪上以最大速度振荡10 min（让Lysis Tube保持水平放置）。若样本数量超过12，延长5-10 min。
例如使用Scientific Industries或Mobio的Vortex-Genie2 涡旋振荡仪。
3. 使用Qiagen的TissueLyser II时，以25Hz研磨10 min。
4. 使用Qiagen的PowerLyzer 24 Homogenizer时，以2000 rpm 的速度均质化30 sec ，暂停30 sec，然后以2000 rpm的速度再次均质化30 sec。
5. 使用MP Biomedicals的FastPrep-24时，推荐速度为 6.0，时间为 40 sec。
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Product Introduction
This kit provides a simple, fast, and efficient method for extracting DNA, suitable for soil and fecal samples. In the presence of high salt, DNA binds to the surface of silica coated Magheads. After rinsing, high-purity DNA is eluted in Buffer EBL or deionized water. The purified DNA has good purity (A260/280 ratio between 1.7-1.9) and high integrity (>15 kb), and can be used for downstream experiments such as second-generation sequencing, quantitative PCR, and chip detection.
This reagent kit can be matched with CWE2100 or CWE3200 32 channel nucleic acid extractors and CWE9600 or CWE960 96 channel nucleic acid extractors for simple and fast high-throughput extraction, greatly reducing the workload of experimenters and human errors in experiments.
Self provided instruments and reagents
1. Constant temperature mixer
2. 2/15 mL magnetic rack
3. 32 channel nucleic acid extractor
4. 96 channel nucleic acid extractor
5. 96 DW Plate
6.8 channel Comb
7. Spin tips pack
8. Anhydrous ethanol, isopropanol
9. Vortex oscillator or tissue grinder
matters needing attention
1. Before the first use, add anhydrous ethanol to Buffer GW1 and Buffer GW2 according to the label of the reagent bottle and mark them properly.
2. Magheads are strictly prohibited from freezing or centrifugation. Freezing and centrifugation may cause irreversible damage to Magheads.
Operation steps
1. Sample pre-processing
1.1 Short centrifugation of Lysis Tube to allow beads to settle at the bottom.
1.2
A. Add 0.1-0.3g of soil or fecal samples to Lysis Tube, and add 740-820 μ L Buffer QSL and 4 μ LRNase A, tighten the tube cover and briefly vortex to mix.
B. If the fecal sample is stored in non cracked fecal preservation solution, add 200 to the Ly - sis Tube μ L-600 μ L solid-liquid mixture, centrifuge at 13000 rpm for 1 minute, discard the storage solution (if the amount of solid after centrifugation is too small, it can be enriched again, but should not exceed 0.3 g). Join 620 μ L Buffer QSL and 4 μ L RNase A, tighten the tube cover and briefly vortex to mix.
1.3 Fix the Lysis Tube in an oscillating grinding device equipped with a 2mL adapter and process it according to the optimized grinding conditions of your equipment (see appendix).
1.4 Shake the Lysis Tube on a constant temperature mixer at 70 ℃ and 1200rpm for 10 minutes. Subsequently, centrifuge at 13000 rpm for 2 minutes to precipitate solid particles. Transfer 540 μ Transfer the supernatant to a new 2 mL centrifuge tube.
1.5 Add 180 μ L Buffer RIL, vortex for 5 seconds, centrifuge at 13000 rpm for 2 minutes. Attention: Remove the buffer RIL before use and store it at 2-8 ℃ immediately after use.
1.6 Manual extraction shall be operated according to Part 2, and the fully automatic nucleic acid extractor shall be operated according to Part 3 or 4.
2. Manual extraction
2.1 Add 160 in sequence to the centrifuge tube μ L Buffer ML, 480 μ Supernatant from step 1.5, 320 μ L isopropanol, 20 μ L Magheads SN, vortex oscillate and mix for 5 seconds, then place the centrifuge tube on a constant temperature mixer at 25 ℃ and 1600 rpm to oscillate and mix for 10 minutes.
Attention: Before adding Magheads SN, vortex oscillation is required for 20 seconds to fully mix it. Magheads SN can be pre mixed with isopropanol according to the volume of the sample and then added.
2.2 Place the centrifuge tube on a magnetic stand and let it stand for 1 minute. After Magheads are completely adsorbed on the side wall of the centrifuge tube, completely discard the solution (keep the centrifuge tube fixed on the magnetic stand).
2.3 Remove the centrifuge tube from the magnetic frame and add 750 μ L Buffer GW1 (please check if anhydrous ethanol has been added before use), vortex point shake for 1 minute or vortex shake for 5 seconds, then place it on a constant temperature mixer at 25 ℃ and 1600 rpm and shake for 2 minutes (ensure that Magheads are in a mixed state during the shaking process). Afterwards, place the centrifuge tube on a magnetic frame and let it stand for 1 minute. After the Magheads are completely adsorbed on the side wall of the centrifuge tube, gently invert the magnetic frame to wash the impurities on the centrifuge tube cover and completely discard the solution (keep the centrifuge tube fixed on the magnetic frame).
2.4 Repeat step 2.3.
2.5 Remove the centrifuge tube from the magnetic frame and add 750 μ L Buffer GW2 (please check if anhydrous ethanol has been added before use), vortex point shake for 1 minute or vortex shake for 5 seconds, and then place it on a constant temperature mixer at 25 ℃ and 1600 rpm to shake and mix for 2 minutes (ensure that Magheads are in a mixed state during the shaking process). Afterwards, place the centrifuge tube on a magnetic frame and let it stand for 1 minute. After the Magheads are completely adsorbed on the side wall of the centrifuge tube, gently invert the magnetic frame to wash the impurities on the centrifuge tube cover and completely discard the solution (keep the centrifuge tube fixed on the magnetic frame).
2.6 Repeat step 2.5.
2.7 Keep the centrifuge tube fixed on the magnetic frame, use a pipette to further remove the solution from the bottom and cover of the centrifuge tube, and then leave it at room temperature for 5-10 minutes to allow the ethanol to evaporate completely (visually observe that the surface of the magnetic beads becomes matte and there are no dry cracks).
2.8 Remove the centrifuge tube from the magnetic frame and add 50-200 μ L Buffer EBL. Vortex oscillation causes the magnetic beads to completely suspend in the eluent and then place them on a constant temperature mixer at 56 ℃ and 1600 rpm for 10 minutes of shaking and elution, or incubate the centrifuge tube in a 56 ℃ water bath for 10 minutes, with vortex oscillation every 3 minutes for 10 seconds.
2.9 Place the centrifuge tube on a magnetic rack and let it stand for 2 minutes. After Magheads are completely adsorbed on the side wall of the centrifuge tube, transfer the eluent to a new centrifuge tube using a pipette and store it at -20 ℃ for later use.
3. Matching with a 32 channel nucleic acid extractor
3.1 Add the corresponding reagents to the 96DW deep well plate according to the table below.
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Note:
1 Add reagents in the 1&7 Columns in order.
2.Before using GW1 and GW2, please check if anhydrous ethanol has been added.
3. Buffer GW2 and Magheads SN can be pre mixed according to the sample size according to the above volume, using pre vortex for 10 seconds to mix well.
3.2 Place the deep well plate and magnetic sleeve with added reagents at the corresponding positions of the 32 channel nucleic acid extractor and run the extraction program. After about 40 minutes, the program ends running.
3.3 Transfer the elution products from columns 6 and 12 of the deep well plate to a 1.5 mL centrifuge tube and store at -20 ℃.
4. Matching with 96 channel nucleic acid extractor
4.1 Add the corresponding reagents to the 96DW deep well plate according to the table below.
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Note:
1 Add reagents in Plate1 in order.
2.Before using GW1 and GW2, please check if anhydrous ethanol has been added.
3. Buffer GW2 and Magheads SN can be pre mixed according to the sample size according to the above volume, and mixed thoroughly by vortex for 10 seconds before use.
4.2 Place the deep well plate and magnetic sleeve that have been added to the reagent in the corresponding positions of CWE960 (magnetic sleeve in position 4) or CWE9600 (magnetic sleeve in position 8), run the extraction program, and after about 50 minutes, the program will end.
4.3 Transfer the elution products from Plate 6 of the deep well plate to a 1.5 mL centrifuge tube and store at -20 ℃.
Appendix:
Grind the sample using one of the following methods:
1. Manually vortex oscillate at maximum speed on the vortex oscillator for 10 minutes.
2. On a vortex oscillator equipped with a 1.5-2 mL horizontal centrifuge tube holder, oscillate at maximum speed for 10 minutes (keeping the Lysis Tube horizontal). If the sample size exceeds 12, extend by 5-10 minutes.
For example, using Scientific Industries or Mobile's Vortex Genie2 vortex oscillator.
3.When using Qiagen's TissueLyser II, grind at 25Hz for 10 minutes.
4.When using Qiagen's PowerLyzer 24 Homogenizer, homogenize at 2000 rpm for 30 seconds, pause for 30 seconds, and then homogenize again at 2000 rpm for 30 seconds.
5.When using FastPrep-24 from MP Biomedicals, the recommended speed is 6.0 and the time is 40 seconds.
规格
英文名称
Magbead Soil And Stool DNA Kit
储存与运输
储存条件
室温
运输条件
常规运输
技术文档
高级数据
质检证书（CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
技术文档和文章
溶液计算器
常见问题
本产品应如何储存？
请于室温条件下保存。
本产品如何运输？
本产品按常温标准条件运输，无需温控包装。
如何获取批次专用COA？
每个订单都会随附分析证书（COA），您也可以从账户订单历史中下载。如需订购前的COA，请联系您的客户代表，或使用PDP上的"申请COA"按钮——我们将提供代表性批次的COA。
试剂盒包含哪些组分？
完整组分列表见PDP和试剂盒说明书。个别组分的替换品可单独订购——请联系您的客户代表。
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
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