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
产品内容
Product Introduction
This kit provides a simple, fast, and efficient method for extracting DNA from paraffin embedded tissues. During the experiment, non-toxic dewaxing agents can be used to remove paraffin and reduce the harm to the experimenter. After tissue lysis, DNA binds to the surface of silicon coated magnetic beads. After rinsing, high-purity DNA is eluted in TE or deionized water. Purified DNA can be directly used for PCR, Real time PCR, SNP genotyping, STR genotyping, second-generation sequencing, and pharmacogenomic research. The yield and fragment size of DNA are closely related to sample type, storage conditions, and storage time.
Self prepared reagent:
1 Fully automatic nucleic acid extractor
2. 96 DW PLate, 8 channel Comb
3.3 Anhydrous ethanol and isopropanol
Preparation and important precautions before the experiment
1. After obtaining the sample, it is necessary to fix it in 4% -10% formalin as soon as possible, with a suitable fixation time of 14-24 hours. Excessive time can lead to genome breakage and affect downstream experiments. If formaldehyde is fixed for too long or the sample is stored for too long (>1 year), it is easy to damage DNA integrity and prevent the expansion of long fragments.
2. Ensure that the sample before embedding is thoroughly dehydrated, as residual formalin will inhibit the action of Protein K.
3.Before the first use, anhydrous ethanol should be added to Buffer GW1 and Buffer GW2 according to the instructions on the reagent bottle label.
4. Before use, please check if there is any crystallization or precipitation in Buffer GTL, Buffer GL, and Buffer DS. If there is any crystallization or precipitation, please dissolve Buffer GTL, Buffer GL, and Buffer DS again in a 56 ℃ water bath.
5. If downstream experiments are sensitive to RNA contamination, 2 can be added before adding Buffer GL μ RNase A of L DNase Free (100 mg/mL). If RNase A in the reagent kit is not used for a long time, it is recommended to store it at -20 ℃.
6. Preheat the water bath or constant temperature mixer to 56 ℃ before the experiment begins.
7. Magheads PN is strictly prohibited from freezing and high-speed centrifugation, otherwise it may cause irreversible damage to Magheads PN. Please shake and mix Magheads PN thoroughly and evenly every time it is used.
Operation steps
1、 Paraffin embedded samples:
1. Use a surgical knife to trim off excess paraffin from the tissue block, expose the tissue, and cut it into 5-10 µ m thin slices.
2. Take approximately 1 x 1 cm ² Slices (approximately 3-8 in total) are placed in centrifuge tubes (provided by oneself) and 160 are added μ L Buffer DS, vortex oscillation for 10 seconds, add 180 more μ L Buffer GTL and 20 μ L Protein K, vortex oscillation for 10 seconds. Centrifuge at 12000 rpm and 25 ℃ for 1 minute.
Note:
1) If the surface of the sample has already been exposed to air, please discard 2-3 pieces that come into contact with the air and do not use them.
2) DS will solidify below 18 ℃, and if it solidifies, it will not affect the following experiments.
3.Incubate at 56 ℃ for 1 hour until the sample is completely dissolved. Incubate at 90 ℃ for 1 hour. After allowing the sample to stand at room temperature for 30 seconds, centrifuge at 25 ℃ and 12000 rpm for 1 minute. Use a pipette to carefully aspirate the lower aqueous phase (approximately 180 degrees) along the tube wall μ L) In the new centrifuge tube, try to avoid inhaling wax liquid and settling at the bottom of the tube as much as possible.
Note:
1) After incubating at 56 ℃, the sample can be placed at room temperature until the temperature of the water or dry bath reaches 90 ℃, and then incubated at 90 ℃.
2) Optional steps: Add 7 μ L UNG (1 U/ μ L) 50 ℃, 5 minutes, without vibration. The purpose of this step is to reduce the low-frequency occurrence of C>T | G>A transitions (artificial mutations) while effectively preserving the true mutations, thereby minimizing the risk of false positives.
4. Optional steps, if the presence of a small amount of RNA will affect downstream experiments, it is recommended to add 2 to the aqueous phase μ L RNase A solution with a concentration of 100 mg/mL, shake well and let stand at room temperature for 2 minutes.
5. Add 20 uL Protein K and incubate at 65 ℃ and 450 rpm for 15 minutes to form Lysate.
2、 Samples in fixed solutions such as formalin:
1. Take approximately 20 mg of the sample, cut it into small pieces, place it in a centrifuge tube, and add 500 μ L 10mM PBS (pH 7.4), vortex oscillation, centrifugation at 12000 rpm (~13400 × g) for 1 minute, discard supernatant, repeat 3 times.
2. Add 180 to the above pipes μ L Buffer GTL, 20 μ L Protein K, vortex oscillation mixing.
3.Incubate at 56 ℃ for 1 hour until the sample is completely dissolved. Incubate at 90 ℃ for 1 hour. After allowing the sample to stand at room temperature for 30 seconds, centrifuge at 25 ℃ and 12000 rpm for 1 minute. Use a pipette to carefully aspirate the lower aqueous phase (approximately 180 degrees) along the tube wall μ L) In the new centrifuge tube, try to avoid inhaling wax liquid and settling at the bottom of the tube as much as possible.
Note:
1) After incubating at 56 ℃, the sample can be placed at room temperature until the temperature of the water or dry bath reaches 90 ℃, and then incubated at 90 ℃.
2) Optional steps: Add 7 μ L UNG (1U/ μ L) 50 ℃, 5 minutes, without vibration. The purpose of this step is to reduce the low-frequency occurrence of C>T | G>A transitions (artificial mutations) while effectively preserving the true mutations, thereby minimizing the risk of false positives.
4. Optional steps, if the presence of a small amount of RNA will affect downstream experiments, it is recommended to add 2 to the aqueous phase μ L RNase A solution with a concentration of 100 mg/mL, shake well and let stand at room temperature for 2 minutes.
5. Add 20 uL Protein K and incubate at 65 ℃ and 450 rpm for 15 minutes to form Lysate.
Manual operation steps:
1. Add 200 to Lysate μ L Buffer GL and vortex oscillation mixing.
2. Add 300 to the centrifuge tube μ L isopropanol and 20 μ Mix the L-shaped magnetic beads with vortex oscillation for 5 seconds, then fix them in a centrifuge tube Shake and combine on a constant temperature mixer at 25 ℃ and 1600 rpm for 10 minutes.
3. Fix the centrifuge tube on a magnetic rack and let it stand for 1 minute, then discard the solution.
4. Add 750 to the centrifuge tube μ L Buffer GW1 (please check if anhydrous ethanol has been added before use), vortex for 5 seconds, and then place it on a constant temperature mixer at 25 ℃ and 1600 rpm for 2 minutes.
5. Fix the centrifuge tube on a magnetic stand and let it stand for 1 minute, then discard the solution.
6. Repeat steps 4-5.
7. Add 750 to the centrifuge tube μ L Buffer GW2 (please check if anhydrous ethanol has been added before use), vortex for 5 seconds, and then place it on a constant temperature mixer at 25 ℃ and 1600 rpm for 2 minutes.
8. Fix the centrifuge tube on a magnetic rack and let it stand for 1 minute, then discard the solution.
9. Repeat steps 7-8.
10. After a brief centrifugation of the centrifuge tube, fix it back on the magnetic frame and use a pipette to remove the solution at the bottom of the tube. Then, open the lid and let it sit at room temperature for 5-10 minutes to allow the ethanol to fully evaporate.
11. Add 50-200 to the centrifuge tube μ The vortex oscillation after L Buffer TE causes the magnetic beads to fully suspend in the eluent, and then the centrifuge tube is fixed on a constant temperature mixing at 56 ℃ and 1600 rpm to shake and wash for 10 minutes.
12. Fix the centrifuge tube on the magnetic frame and let it stand for 2 minutes, then transfer the eluent to a new centrifuge tube and store it at -20 ℃ for later use.
Matching with fully automatic nucleic acid extractor
1. Add reagents to the 96 DW deep well plate according to the following table:
Note: In columns 1&7, Magheads PN can be mixed with isopropanol in the ratio shown in the table before being added.
2. Place the magnetic sleeve and 96 DW deep well plate into the fully automatic nucleic acid extractor and run the "FFPE DNA program".
3. After about 30 minutes, the program runs and the 96DW deep well plate and magnetic sleeve are removed from the instrument. The elution products from "6&12 Columns" are transferred to a centrifuge tube and stored at -20 ℃ for later use.
Attachment: FFPE DNA Program
|
产品简介
该试剂盒提供了一种简单、快速、高效的从石蜡包埋组织中提取DNA的方法。实验 过程中可采取无毒的脱蜡剂去除石蜡，降低实验过程中对实验者的危害。组织裂解后， DNA结合于硅基包被磁珠表面。漂洗后，高纯度的DNA洗脱于TE或去离子水中。经过 纯化的DNA可以直接用于PCR、 Real-time PCR、 SNP基因分型、STR基因分型、二 代测序和药物基因组学研究等。 DNA的得率、片段大小与样品类型、储存条件及储存时 间有很大关系。
自备试剂：
1. 全自动核酸提取仪
2. 96 DW PLate、 8 channel Comb
3. 3. 无水乙醇、异丙醇
实验前准备及重要注意事项
1. 获得样品后，要尽快将样品在4%-10%的福尔马林中固定，固定时间以14-24小时为 宜，时间过长易导致基因组断裂，影响下游实验。如果甲醛固定时间过长或样本存 放时间过久（＞1年）则易导致DNA完整性受损，无法扩出长片段。
2. 确保包埋前的样品彻底脱水，残留的福尔马林会抑制Proteinase K的作用。
3. 第一次使用前应按试剂瓶标签说明先在Buffer GW1和Buffer GW2中加入无水乙醇。
4. 使用前请检查Buffer GTL、Buffer GL和Buffer DS是否出现结晶或者沉淀，如有结晶 或者沉淀， 请将Buffer GTL、Buffer GL和Buffer DS于56℃水浴重新溶解。
5. 如果下游实验对RNA污染比较敏感，可以在加入Buffer GL前加入2 μL DNase-Free的 RNase A（100 mg/mL），试剂盒中的RNase A如果长时间不用，建议-20℃保存。
6. 实验开始前将水浴锅或恒温混匀仪预热至56℃。
7. Magbeads PN严禁冰冻和高速离心，否则可能会对Magbeads PN造成不可逆的损 害。 Magbeads PN每次使用时请充分振荡混合均匀。
操作步骤
一、石蜡包埋样本：
1. 用手术刀将组织块中多余的石蜡修剪掉，露出组织后切成5-10 µm的薄片。
2. 取约1×1cm²的切片（共约3-8片切片）置于离心管（自备）中，加入160 μL Buffer DS，涡旋震荡10秒，再加入180 μL Buffer GTL和20 μL Proteinase K，涡旋震荡10 秒。 12000 rpm ，25℃ 离心1分钟。
注意：1 ) 如果样品表面已经暴露在空气中，请将接触空气的2-3片弃掉不用。
2）DS低于18℃会凝固，如果凝固不影响下面的实验。
3. 56℃孵育1小时，直至样品完全溶解。 90℃孵育1小时。样品室温静置30秒后25℃、 12000 rpm离心1分钟，用移液器沿管壁小心吸取下层水相（约180μL）于新离心管 中，尽量避免吸入蜡液和管底沉淀。
注意：1）56℃孵育后的样品可置于室温，直至水浴锅或干浴锅温度达到90℃后再把样品置于90℃孵 育。
2）可选步骤：加入7 μL UNG（1 U/μL），50℃,5min ，不震荡。此步骤的目的是降低低频发生的 C>T | G>A转换（人为突变），同时有效保留真实发生的突变，从而使假阳性风险降至最低。
4. 可选步骤，如果少量RNA的存在会对下游实验造成影响，建议向水相中加入2 μL 浓 度为100 mg/mL的RNase A溶液，震荡混匀，室温放置2 min。
5. 加入20 uL Proteinase K ，65℃, 450 rpm孵育15 min ，形成Lysate。
二、福尔马林等固定液中的样本：
1. 取约20mg的样本，切成小块，置于离心管中，加入500 μL 10mM PBS（PH 7.4）， 涡旋振荡， 12,000 rpm（～13,400×g）离心1分钟，弃上清，重复3次。
2. 上述管中加入180 μL Buffer GTL ， 20 μL Proteinase K，涡旋震荡混匀。
3. 56℃孵育1小时，直至样品完全溶解。 90℃孵育1小时。样品室温静置30秒后25℃、 12000rpm离心1分钟，用移液器沿管壁小心吸取下层水相（约180μL）于新离心管 中，尽量避免吸入蜡液和管底沉淀。
注意：1）56℃孵育后的样品可置于室温，直至水浴锅或干浴锅温度达到90℃后再把样品置于90℃ 孵育。
2）可选步骤：加入7 μL UNG（1U/μL），50℃, 5min ，不震荡。此步骤的目的是 降低低频发生的 C>T | G>A 转换（人为突变），同时有效保留真实发生的突变，从 而使假阳性风险降至最低。
4. 可选步骤，如果少量RNA的存在会对下游实验造成影响，建议向水相中加入2 μL浓 度为100 mg/mL的RNase A溶液，震荡混匀，室温放置2 min。
5. 加入20 uL Proteinase K ，65℃, 450 rpm孵育15 min ，形成Lysate。
手动操作步骤：
1. 向Lysate 中加入200 μL Buffer GL并涡旋震荡混匀。
2. 向离心管中加入300 μL异丙醇与20 μL磁珠后涡旋震荡混匀5秒钟，之后将离心管固
定于25℃、 1600 rpm的恒温混匀仪上震荡结合10分钟。
3. 将离心管固定于磁力架上静置1分钟，之后弃去溶液。
4. 向离心管中加入750 μL Buffer GW1（使用前请检查是否已加入无水乙醇），涡旋震 荡5秒钟后放于25℃、 1600 rpm的恒温混匀仪上震荡结合2分钟。
5. 将离心管固定于磁力架上静置1分钟，之后弃去溶液。
6. 重复步骤4-5。
7. 向离心管中加入750 μL Buffer GW2（使用前请检查是否已加入无水乙醇），涡旋震 荡5秒钟后放于25℃、 1600 rpm的恒温混匀仪上震荡结合2分钟。
8. 将离心管固定于磁力架上静置1分钟，之后弃去溶液。
9. 重复步骤7-8。
10. 离心管短暂离心后，将其重新固定于磁力架上用移液器去除管底溶液，之后开盖室 温放置5-10分钟使乙醇充分挥发。
11. 向离心管中加入50-200 μL Buffer TE后涡旋震荡使磁珠充分悬浮于洗脱液中，之后 将离心管固定于56℃、 1600 rpm的恒温混匀上震荡洗脱10分钟。
12. 将离心管固定于磁力架上静置2分钟，之后将洗脱液转移至新的离心管中-20℃保存 备用。
与全自动核酸提取仪匹配
1. 按下表向96 DW深孔板中加入试剂：
|
注意：在1&7列中，可以先将Magbeads PN与异丙醇按表中比例混匀后再加入。
2. 将磁套与96 DW深孔板放入全自动核酸提取仪中，运行“FFPE DNA程序”。
3. 约30分钟后程序运行结束，将96DW深孔板和磁套从仪器中取出，把“6&12 Colume” 中的洗脱产物转移至离心管中-20℃保存备用。
附： FFPE DNA程序
|
|
Product Introduction
This kit provides a simple, fast, and efficient method for extracting DNA from paraffin embedded tissues. During the experiment, non-toxic dewaxing agents can be used to remove paraffin and reduce the harm to the experimenter. After tissue lysis, DNA binds to the surface of silicon coated magnetic beads. After rinsing, high-purity DNA is eluted in TE or deionized water. Purified DNA can be directly used for PCR, Real time PCR, SNP genotyping, STR genotyping, second-generation sequencing, and pharmacogenomic research. The yield and fragment size of DNA are closely related to sample type, storage conditions, and storage time.
Self prepared reagent:
1 Fully automatic nucleic acid extractor
2. 96 DW PLate, 8 channel Comb
3.3 Anhydrous ethanol and isopropanol
Preparation and important precautions before the experiment
1. After obtaining the sample, it is necessary to fix it in 4% -10% formalin as soon as possible, with a suitable fixation time of 14-24 hours. Excessive time can lead to genome breakage and affect downstream experiments. If formaldehyde is fixed for too long or the sample is stored for too long (>1 year), it is easy to damage DNA integrity and prevent the expansion of long fragments.
2. Ensure that the sample before embedding is thoroughly dehydrated, as residual formalin will inhibit the action of Protein K.
3.Before the first use, anhydrous ethanol should be added to Buffer GW1 and Buffer GW2 according to the instructions on the reagent bottle label.
4. Before use, please check if there is any crystallization or precipitation in Buffer GTL, Buffer GL, and Buffer DS. If there is any crystallization or precipitation, please dissolve Buffer GTL, Buffer GL, and Buffer DS again in a 56 ℃ water bath.
5. If downstream experiments are sensitive to RNA contamination, 2 can be added before adding Buffer GL μ RNase A of L DNase Free (100 mg/mL). If RNase A in the reagent kit is not used for a long time, it is recommended to store it at -20 ℃.
6. Preheat the water bath or constant temperature mixer to 56 ℃ before the experiment begins.
7. Magheads PN is strictly prohibited from freezing and high-speed centrifugation, otherwise it may cause irreversible damage to Magheads PN. Please shake and mix Magheads PN thoroughly and evenly every time it is used.
Operation steps
1、 Paraffin embedded samples:
1. Use a surgical knife to trim off excess paraffin from the tissue block, expose the tissue, and cut it into 5-10 µ m thin slices.
2. Take approximately 1 x 1 cm ² Slices (approximately 3-8 in total) are placed in centrifuge tubes (provided by oneself) and 160 are added μ L Buffer DS, vortex oscillation for 10 seconds, add 180 more μ L Buffer GTL and 20 μ L Protein K, vortex oscillation for 10 seconds. Centrifuge at 12000 rpm and 25 ℃ for 1 minute.
Note:
1) If the surface of the sample has already been exposed to air, please discard 2-3 pieces that come into contact with the air and do not use them.
2) DS will solidify below 18 ℃, and if it solidifies, it will not affect the following experiments.
3.Incubate at 56 ℃ for 1 hour until the sample is completely dissolved. Incubate at 90 ℃ for 1 hour. After allowing the sample to stand at room temperature for 30 seconds, centrifuge at 25 ℃ and 12000 rpm for 1 minute. Use a pipette to carefully aspirate the lower aqueous phase (approximately 180 degrees) along the tube wall μ L) In the new centrifuge tube, try to avoid inhaling wax liquid and settling at the bottom of the tube as much as possible.
Note:
1) After incubating at 56 ℃, the sample can be placed at room temperature until the temperature of the water or dry bath reaches 90 ℃, and then incubated at 90 ℃.
2) Optional steps: Add 7 μ L UNG (1 U/ μ L) 50 ℃, 5 minutes, without vibration. The purpose of this step is to reduce the low-frequency occurrence of C>T | G>A transitions (artificial mutations) while effectively preserving the true mutations, thereby minimizing the risk of false positives.
4. Optional steps, if the presence of a small amount of RNA will affect downstream experiments, it is recommended to add 2 to the aqueous phase μ L RNase A solution with a concentration of 100 mg/mL, shake well and let stand at room temperature for 2 minutes.
5. Add 20 uL Protein K and incubate at 65 ℃ and 450 rpm for 15 minutes to form Lysate.
2、 Samples in fixed solutions such as formalin:
1. Take approximately 20 mg of the sample, cut it into small pieces, place it in a centrifuge tube, and add 500 μ L 10mM PBS (pH 7.4), vortex oscillation, centrifugation at 12000 rpm (~13400 × g) for 1 minute, discard supernatant, repeat 3 times.
2. Add 180 to the above pipes μ L Buffer GTL, 20 μ L Protein K, vortex oscillation mixing.
3.Incubate at 56 ℃ for 1 hour until the sample is completely dissolved. Incubate at 90 ℃ for 1 hour. After allowing the sample to stand at room temperature for 30 seconds, centrifuge at 25 ℃ and 12000 rpm for 1 minute. Use a pipette to carefully aspirate the lower aqueous phase (approximately 180 degrees) along the tube wall μ L) In the new centrifuge tube, try to avoid inhaling wax liquid and settling at the bottom of the tube as much as possible.
Note:
1) After incubating at 56 ℃, the sample can be placed at room temperature until the temperature of the water or dry bath reaches 90 ℃, and then incubated at 90 ℃.
2) Optional steps: Add 7 μ L UNG (1U/ μ L) 50 ℃, 5 minutes, without vibration. The purpose of this step is to reduce the low-frequency occurrence of C>T | G>A transitions (artificial mutations) while effectively preserving the true mutations, thereby minimizing the risk of false positives.
4. Optional steps, if the presence of a small amount of RNA will affect downstream experiments, it is recommended to add 2 to the aqueous phase μ L RNase A solution with a concentration of 100 mg/mL, shake well and let stand at room temperature for 2 minutes.
5. Add 20 uL Protein K and incubate at 65 ℃ and 450 rpm for 15 minutes to form Lysate.
Manual operation steps:
1. Add 200 to Lysate μ L Buffer GL and vortex oscillation mixing.
2. Add 300 to the centrifuge tube μ L isopropanol and 20 μ Mix the L-shaped magnetic beads with vortex oscillation for 5 seconds, then fix them in a centrifuge tube Shake and combine on a constant temperature mixer at 25 ℃ and 1600 rpm for 10 minutes.
3. Fix the centrifuge tube on a magnetic rack and let it stand for 1 minute, then discard the solution.
4. Add 750 to the centrifuge tube μ L Buffer GW1 (please check if anhydrous ethanol has been added before use), vortex for 5 seconds, and then place it on a constant temperature mixer at 25 ℃ and 1600 rpm for 2 minutes.
5. Fix the centrifuge tube on a magnetic stand and let it stand for 1 minute, then discard the solution.
6. Repeat steps 4-5.
7. Add 750 to the centrifuge tube μ L Buffer GW2 (please check if anhydrous ethanol has been added before use), vortex for 5 seconds, and then place it on a constant temperature mixer at 25 ℃ and 1600 rpm for 2 minutes.
8. Fix the centrifuge tube on a magnetic rack and let it stand for 1 minute, then discard the solution.
9. Repeat steps 7-8.
10. After a brief centrifugation of the centrifuge tube, fix it back on the magnetic frame and use a pipette to remove the solution at the bottom of the tube. Then, open the lid and let it sit at room temperature for 5-10 minutes to allow the ethanol to fully evaporate.
11. Add 50-200 to the centrifuge tube μ The vortex oscillation after L Buffer TE causes the magnetic beads to fully suspend in the eluent, and then the centrifuge tube is fixed on a constant temperature mixing at 56 ℃ and 1600 rpm to shake and wash for 10 minutes.
12. Fix the centrifuge tube on the magnetic frame and let it stand for 2 minutes, then transfer the eluent to a new centrifuge tube and store it at -20 ℃ for later use.
Matching with fully automatic nucleic acid extractor
1. Add reagents to the 96 DW deep well plate according to the following table:
|
2. Place the magnetic sleeve and 96 DW deep well plate into the fully automatic nucleic acid extractor and run the "FFPE DNA program".
3. After about 30 minutes, the program runs and the 96DW deep well plate and magnetic sleeve are removed from the instrument. The elution products from "6&12 Columns" are transferred to a centrifuge tube and stored at -20 ℃ for later use.
Attachment: FFPE DNA Program
|
规格
英文名称
NuClean Magbead FFPE 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
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。
Copyright © 2023至今 上海阿拉丁生化科技股份有限公司 保留所有权利。
危险品化学品经营许可证（带存储）