本试剂盒为微量样本基因组DNA的提取提供了一个简单快速的解决方案，该方法采用超顺磁性的磁性粒子纯化技术，提取过程中无需使用有毒的酚氯仿抽提，操作简单方便，纯化的DNA产物可直接用作PCR模板、酶切、杂交等分子生物学实验。其处理后的产物用于科研检测使用。
| 产品号 | 组件名称 | 物理外观 | 48T | 储存温度 | | M1522612A | 裂解液AL | 液体 | 70 mL | RT. | | M1522612B | 裂解液ML | 液体 | 14 mL | RT. | | M1522612C | 蛋白酶K | 液体 | 1.1 mL | 2-8℃. | | M1522612D | 裂解结合液RLB | 液体 | 35 mL | RT. | | M1522612E | 磁珠MXVB | 液体 | 1.1 mL | 2-8℃. | | M1522612F | 洗涤液MW1 | 液体 | 40 mL | RT. | | M1522612G | 洗涤液BW1 | 液体 | 40 mL | RT. | | M1522612H | 洗涤液DW2 | 液体 | 40 mL | RT. | | M1522612I | 洗脱液DE | 液体 | 5 mL | RT. |
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储存条件
M1522612C, M1522612E：2-8℃保存，12个月。
M1522612A, M1522612B, M1522612D, M1522612F, M1522612G, M1522612H, M1522612I：室温保存，12个月。
适配仪器
手工提取
注意事项
1. 使用前请仔细阅读本说明书。
2. 本试剂盒中的多种缓冲液含有刺激性的胍盐，务必戴上手套，并按照安全标准预防措施来处理。不要让缓冲液接触到皮肤、眼睛以及黏膜，如果确实发生，请立即用大量清水清洗并就医。
3. 如因气温较低，溶液出现沉淀，需要30℃水浴至沉淀完全溶解后方可使用。
4. 如因气温较低，裂解液AL、裂解液ML出现沉淀，可60℃水浴至沉淀完全溶解后使用。
5.如因气温较低，用力振荡后磁珠无法重悬，请勿使用。
6. 本试剂盒中的多种缓冲液含胍盐，请勿用氧化性消毒剂如次氯酸钠进行处理，否则会释放有毒气体，须按医疗废物进行处理。
7. 本试剂盒需配套使用，不同批号试剂盒中各组分不可以互换或混用。
使用方法
1. 样本要求
- 适用标本类型：微量血、微量组织、骨髓、干血斑、唾液斑、漱口水、口香糖、牙刷、烟蒂、指甲、毛发、虾皮等微量样本。
- 标本保存和运输：标本可立即用于测试，也可以保存于-70℃或更低温度待测，保存期为6个月，应避免反复冻融。标本运输采用冷链运输。
- 冻融要求：速冻速融，避免反复冻融。
2. 自备试验器材
- 恒温水浴锅（当溶液出现沉淀时使用）、干式恒温仪
- 磁力架（1.5/2.0 mL）、1.5/2.0/50.0 mL无菌无核酸酶离心管、各种规格移液器及Tips（20、200、1000 μL）
- 1 M DTT（提取指甲、毛发、骨髓、虾皮时使用）
3. 试剂准备
4. 操作步骤
磁分离操作建议（步骤4.5开始）：离心管置于磁力架后，轻轻地左右旋转，待磁珠聚集于贴近磁力架的管壁后，轻轻地颠倒磁力架数次，使管盖上的磁珠也聚集到管壁，静置数分钟至溶液澄清即可（静置时长视磁力架磁性而定）。
4.1 根据不同样本类型选择对应步骤进行处理
- 微量血：取1~100 μL血液到1.5 mL的离心管中，加入200 μL裂解液ML和20 μL蛋白酶K，涡旋混匀，56℃温浴10分钟，温浴期间每3分钟涡旋混匀15秒。
- 微量组织：取不超过10 mg的组织到1.5 mL的离心管中，加入200 μL裂解液AL和20 μL蛋白酶K，涡旋混匀，56℃温浴直至样本充分消化，时间大约40分钟到1小时，可使用研磨器研磨组织以促进组织消化，温浴期间每10分钟涡旋混匀15秒。
- 骨髓：取不超过10 mg的骨髓到1.5 mL的离心管中，加入200 μL裂解液AL，20 μL蛋白酶K，20 μL DTT(1 M)，涡旋混匀，56℃温浴直至样本充分消化，时间大约40分钟到1小时，可使用研磨器研磨组织以促进组织消化，温浴期间每10分钟涡旋混匀15秒。
- 干血斑：取一个干血点剪碎到1.5 mL的离心管中，加入300 μL裂解液AL和20 μL蛋白酶K，涡旋混匀，56~75℃温浴40分钟，温浴期间10分钟涡旋混匀15秒。
- 唾液斑：取一个唾液点剪碎到1.5 mL的离心管中，加入300 μL裂解液AL和20 μL蛋白酶K，涡旋混匀，56℃温浴40分钟，温浴期间10分钟涡旋混匀15秒。
- 漱口水：在50 mL离心管中添加10~20 mL漱口水样本，1800×g离心5分钟，将上清小心倒弃。向沉淀中加入200 μL裂解液ML重悬，将全部悬液转移至1.5 mL的离心管中。加入20 μL蛋白酶K，涡旋混匀，56℃温浴40分钟，温浴期间每10分钟涡旋混匀15秒。
- 口香糖：取约0.4 g咀嚼过的口香糖剪碎至2.0 mL离心管中，加入500 μL裂解液AL和20 μL蛋白酶K，涡旋混匀，56℃温浴40分钟，温浴期间每10分钟涡旋混匀15秒。（注意：尽可能使口香糖浸没在裂解液AL中）
- 牙刷：剪取或切取牙刷毛至2.0 mL离心管中，加入500~700 μL裂解液AL和20 μL蛋白酶K，涡旋混匀，56℃温浴40分钟，温浴期间每10分钟涡旋混匀15秒。（注意：尽可能使牙刷毛浸没在裂解液AL中；干燥牙刷推荐700 μL裂解液AL，湿润牙刷毛推荐500 μL）
- 烟蒂：取一个烟蒂至2.0 mL离心管中，加入1000~1200 μL裂解液AL和20 μL蛋白酶K，涡旋混匀，56℃温浴40分钟~1小时，温浴期间每10分钟涡旋混匀15秒。（注意：尽可能使裂解液AL湿润烟蒂）
- 指甲：取5 mg指甲剪碎到1.5 mL离心管中，加入250 μL裂解液ML，20 μL蛋白酶K，20 μL DTT (1 M)，涡旋混匀，56℃温浴1小时，温浴期间每10分钟涡旋混匀15秒。
- 未染色毛发：取10 mg长度为1 cm的毛发到1.5 mL离心管中，加入250 μL裂解液ML，20 μL蛋白酶K，20 μl DTT (1 M)，涡旋混匀，56℃温浴1小时，可延长消化时间使毛发完全消化，温浴期间每15分钟涡旋混匀30秒。
- 染色毛发（需额外购买我司沉淀液SPS，并按以下流程操作）：取10 mg长度为1 cm的毛发到1.5 mL离心管中，加入200 μL裂解液ML，20 μL蛋白酶K，20 μL DTT (1 M)，涡旋混匀，56℃振荡温浴1小时（如恒温仪无振荡功能，可每15分钟涡旋30秒促进裂解）。加入200 μL沉淀液SPS混匀后，4℃冰浴10分钟，10,000×g离心3分钟。
- 虾皮：取10 mg虾皮剪碎到1.5 mL离心管中，加入250 μL裂解液AL，20 μL蛋白酶K，20 μL DTT (1 M)，涡旋混匀，56℃温浴1小时，温浴期间每10分钟涡旋混匀15秒。
4.2 样本消化后短暂离心以收集管壁及管盖的液体。
4.3 转移温浴后消化液至新的1.5 mL离心管中，不同样本转移的消化液量如下。
- 微量血：全部消化液。
- 微量组织：全部消化液。
- 骨髓：全部消化液。
- 干血斑：全部消化液。
- 唾液斑：全部消化液。
- 漱口水：全部消化液。
- 口香糖：300 μL消化液，切勿吸到口香糖。
- 牙刷：300 μL消化液，切勿吸到牙刷毛。
- 烟蒂：300 μL消化液。
- 指甲：全部消化液。
- 未染色毛发：全部消化液。
- 染色毛发：200~300 μL上清液。
- 虾皮：全部消化液。
4.4 加入600 μL裂解结合液RLB，20 µL磁珠MXVB，振荡或涡旋5~7分钟使磁珠吸附DNA。
4.5 转至磁力架上进行磁分离至溶液澄清，吸弃溶液。
4.6 加入700 µL洗涤液MW1，涡旋10秒后进行磁分离至溶液澄清，吸弃溶液。
4.7 加入700 µL洗涤液BW1，涡旋10秒后进行磁分离至溶液澄清，吸弃溶液。
4.8 加入700 µL洗涤液DW2，涡旋10秒后进行磁分离至溶液澄清，吸弃溶液。
4.9 掌上离心机瞬时离心后，转至磁力架上吸附至溶液澄清，充分吸弃所有溶液，打开管盖，空气干燥3~5分钟。
注意：乙醇残留会抑制后续的酶反应，所以晾干时要确保乙醇挥发干净。切勿干燥过久，以免影响后续洗脱效果。
4.10 加入30~50 µL洗脱液DE，高速涡旋2~3分钟打散磁珠。
4.11 60℃温浴5分钟，然后高速涡旋30秒。
4.12 瞬时离心收集管盖液滴至管中，转移至磁力架上进行磁分离。
4.13 把上清基因组DNA转移至新的离心管中待用。如果不立即使用，请存储于-15~-25℃，长期保存请放置于-70℃或更低的温度。
方法的局限性
1. 对于某些取样不佳的样本，提取产物中可能存在少量杂质抑制PCR扩增。
2. 样本核酸的提取效果与样本收集、处理、运输和保存质量密切相关，其中任何失误都将会导致结果不准确。
This kit provides a simple and rapid solution for genomic DNA extraction from trace samples. The method employs superparamagnetic bead purification technology, eliminating the need for toxic phenol‑chloroform extraction. The procedure is straightforward and convenient, and the purified DNA can be used directly as a template for PCR, restriction enzyme digestion, hybridization, and other molecular biology experiments. The purified DNA is intended for research use only.
| Product Code | Component | Appearance | 48T | Storage | | M1522612A | Lysis Buffer AL | Liquid | 70 mL | RT | | M1522612B | Lysis Buffer ML | Liquid | 14 mL | RT | | M1522612C | Proteinase K | Liquid | 1.1 mL | 2–8 °C | | M1522612D | Lysis/Binding Buffer RLB | Liquid | 35 mL | RT | | M1522612E | Magnetic Beads MXVB | Liquid | 1.1 mL | 2–8 °C | | M1522612F | Wash Buffer MW1 | Liquid | 40 mL | RT | | M1522612G | Wash Buffer BW1 | Liquid | 40 mL | RT | | M1522612H | Wash Buffer DW2 | Liquid | 40 mL | RT | | M1522612I | Elution Buffer DE | Liquid | 5 mL | RT |
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Storage Conditions
M1522612C, M1522612E: Store at 2–8 °C for 12 months.
M1522612A, M1522612B, M1522612D, M1522612F, M1522612G, M1522612H, M1522612I: Store at room temperature for 12 months.
Compatible Instrumentation
Manual extraction.
Precautions
Read the entire manual carefully before use.
Several buffers contain irritating guanidine salts. Always wear gloves and follow standard safety precautions. Avoid contact with skin, eyes, and mucous membranes. If contact occurs, rinse immediately with plenty of water and seek medical attention.
If precipitates appear due to low temperature, warm the solution at 30 °C until precipitates are completely dissolved before use.
If precipitates appear in Lysis Buffer AL or Lysis Buffer ML due to low temperature, warm at 60 °C until completely dissolved before use.
If magnetic beads cannot be resuspended by vigorous shaking due to low temperature, do not use.
Several buffers contain guanidine salts. Do not treat with oxidizing disinfectants (e.g., sodium hypochlorite) as this may release toxic gas. Dispose as medical waste.
This kit is intended for use as an integrated system. Do not interchange or mix components from different lot numbers.
Protocol
1. Sample Requirements
Applicable sample types: Trace blood, trace tissue, bone marrow, dried blood spots, saliva stains, mouthwash, chewing gum, toothbrushes, cigarette butts, fingernails, hair, shrimp shells, and other trace samples.
Sample preservation and transport: Samples can be processed immediately or stored at –70 °C or lower for up to 6 months. Avoid repeated freeze‑thaw cycles. Transport samples under cold chain conditions.
Freeze‑thaw requirements: Quick freeze and quick thaw; avoid repeated freeze‑thaw cycles.
2. Materials to Be Prepared by User
Water bath (for dissolving precipitates), dry thermostat bath
Magnetic stand (for 1.5/2.0 mL tubes)
1.5/2.0/50.0 mL sterile, nuclease‑free centrifuge tubes
Various pipettes and tips (20 µL, 200 µL, 1000 µL)
1 M DTT (for fingernails, hair, bone marrow, shrimp shells)
3. Reagent Preparation
Check solutions for precipitates and ensure magnetic beads can be resuspended.
4. Procedure
Recommended magnetic separation procedure (starting from step 4.5): Place the tube on the magnetic stand, gently rotate left and right until the beads aggregate on the tube wall closest to the magnet. Gently invert the magnetic stand several times to collect beads from the cap onto the wall. Let stand for several minutes until the solution is clear (stand time depends on the magnet strength).
4.1 Sample processing according to sample type
Trace blood: Transfer 1–100 μL of blood to a 1.5 mL centrifuge tube. Add 200 μL of Lysis Buffer ML and 20 μL of Proteinase K. Vortex to mix. Incubate at 56 °C for 10 minutes, vortexing for 15 seconds every 3 minutes during incubation.
Trace tissue: Transfer up to 10 mg of tissue to a 1.5 mL centrifuge tube. Add 200 μL of Lysis Buffer AL and 20 μL of Proteinase K. Vortex to mix. Incubate at 56 °C until the sample is fully digested (approximately 40 minutes to 1 hour). A homogenizer may be used to aid tissue digestion. Vortex for 15 seconds every 10 minutes during incubation.
Bone marrow: Transfer up to 10 mg of bone marrow to a 1.5 mL centrifuge tube. Add 200 μL of Lysis Buffer AL, 20 μL of Proteinase K, and 20 μL of 1 M DTT. Vortex to mix. Incubate at 56 °C until fully digested (approximately 40 minutes to 1 hour). A homogenizer may be used. Vortex for 15 seconds every 10 minutes during incubation.
Dried blood spot: Cut one dried blood spot into small pieces and transfer to a 1.5 mL centrifuge tube. Add 300 μL of Lysis Buffer AL and 20 μL of Proteinase K. Vortex to mix. Incubate at 56–75 °C for 40 minutes, vortexing for 15 seconds every 10 minutes.
Saliva stain: Cut one saliva stain into small pieces and transfer to a 1.5 mL centrifuge tube. Add 300 μL of Lysis Buffer AL and 20 μL of Proteinase K. Vortex to mix. Incubate at 56 °C for 40 minutes, vortexing for 15 seconds every 10 minutes.
Mouthwash: Transfer 10–20 mL of mouthwash to a 50 mL centrifuge tube. Centrifuge at 1800×g for 5 minutes. Carefully discard the supernatant. Resuspend the pellet in 200 μL of Lysis Buffer ML and transfer the entire suspension to a 1.5 mL centrifuge tube. Add 20 μL of Proteinase K, vortex to mix, and incubate at 56 °C for 40 minutes, vortexing for 15 seconds every 10 minutes.
Chewing gum: Take approximately 0.4 g of chewed gum, cut into small pieces, and transfer to a 2.0 mL centrifuge tube. Add 500 μL of Lysis Buffer AL and 20 μL of Proteinase K. Vortex to mix. Incubate at 56 °C for 40 minutes, vortexing for 15 seconds every 10 minutes. (Note: Ensure the gum is submerged as much as possible in Lysis Buffer AL.)
Toothbrush: Cut or snip toothbrush bristles into a 2.0 mL centrifuge tube. Add 500–700 μL of Lysis Buffer AL and 20 μL of Proteinase K. Vortex to mix. Incubate at 56 °C for 40 minutes, vortexing for 15 seconds every 10 minutes. (Note: Submerge bristles as much as possible; use 700 μL for dry brushes, 500 μL for wet bristles.)
Cigarette butt: Place one cigarette butt into a 2.0 mL centrifuge tube. Add 1000–1200 μL of Lysis Buffer AL and 20 μL of Proteinase K. Vortex to mix. Incubate at 56 °C for 40 minutes to 1 hour, vortexing for 15 seconds every 10 minutes. (Note: Ensure the butt is thoroughly wetted by the lysis buffer.)
Fingernail: Cut 5 mg of fingernail into small pieces and transfer to a 1.5 mL centrifuge tube. Add 250 μL of Lysis Buffer ML, 20 μL of Proteinase K, and 20 μL of 1 M DTT. Vortex to mix. Incubate at 56 °C for 1 hour, vortexing for 15 seconds every 10 minutes.
Unstained hair: Transfer 10 mg of hair (1 cm pieces) to a 1.5 mL centrifuge tube. Add 250 μL of Lysis Buffer ML, 20 μL of Proteinase K, and 20 μL of 1 M DTT. Vortex to mix. Incubate at 56 °C for 1 hour (digestion time can be extended until hair is fully digested), vortexing for 30 seconds every 15 minutes.
Stained hair (requires separate purchase of our Precipitation Solution SPS; follow the procedure below): Transfer 10 mg of stained hair (1 cm pieces) to a 1.5 mL centrifuge tube. Add 200 μL of Lysis Buffer ML, 20 μL of Proteinase K, and 20 μL of 1 M DTT. Vortex to mix. Incubate at 56 °C with shaking for 1 hour (if the incubator lacks a shaking function, vortex for 30 seconds every 15 minutes to promote lysis). Add 200 μL of Precipitation Solution SPS, mix well, then chill on ice for 10 minutes. Centrifuge at 10,000×g for 3 minutes.
Shrimp shell: Cut 10 mg of shrimp shell into small pieces and transfer to a 1.5 mL centrifuge tube. Add 250 μL of Lysis Buffer AL, 20 μL of Proteinase K, and 20 μL of 1 M DTT. Vortex to mix. Incubate at 56 °C for 1 hour, vortexing for 15 seconds every 10 minutes.
4.2 After digestion, briefly centrifuge to collect liquid from the walls and cap.
4.3 Transfer the digested lysate to a new 1.5 mL centrifuge tube. The volume to transfer varies by sample type:
Trace blood: entire lysate
Trace tissue: entire lysate
Bone marrow: entire lysate
Dried blood spot: entire lysate
Saliva stain: entire lysate
Mouthwash: entire lysate
Chewing gum: 300 μL lysate, avoid taking any gum pieces
Toothbrush: 300 μL lysate, avoid taking bristles
Cigarette butt: 300 μL lysate
Fingernail: entire lysate
Unstained hair: entire lysate
Stained hair: 200–300 μL supernatant
Shrimp shell: entire lysate
4.4 Add 600 μL of Lysis/Binding Buffer RLB and 20 μL of Magnetic Beads MXVB. Shake or vortex for 5–7 minutes to allow the beads to bind DNA.
4.5 Place the tube on a magnetic stand for magnetic separation until the solution is clear. Aspirate and discard the supernatant.
4.6 Add 700 μL of Wash Buffer MW1. Vortex for 10 seconds, then separate magnetically until clear. Aspirate and discard the supernatant.
4.7 Add 700 μL of Wash Buffer BW1. Vortex for 10 seconds, then separate magnetically until clear. Aspirate and discard the supernatant.
4.8 Add 700 μL of Wash Buffer DW2. Vortex for 10 seconds, then separate magnetically until clear. Aspirate and discard the supernatant.
4.9 Briefly centrifuge in a microcentrifuge. Place the tube on the magnetic stand and allow the solution to become clear. Aspirate and discard all remaining liquid thoroughly. Open the cap and air‑dry for 3–5 minutes.
Note: Residual ethanol will inhibit downstream enzymatic reactions. Ensure that ethanol is completely evaporated during drying. Do not over‑dry, as this may affect elution efficiency.
4.10 Add 30–50 μL of Elution Buffer DE. Vortex vigorously for 2–3 minutes to fully resuspend the beads.
4.11 Incubate at 60 °C for 5 minutes, then vortex vigorously for 30 seconds.
4.12 Briefly centrifuge to collect droplets from the cap. Return the tube to the magnetic stand for magnetic separation.
4.13 Transfer the supernatant genomic DNA to a new centrifuge tube for immediate use. If not used immediately, store at –15 to –25 °C. For long‑term storage, keep at –70 °C or lower.
Limitations of the Method
For certain poorly collected samples, the extracted product may contain trace impurities that inhibit PCR amplification.
The efficiency of nucleic acid extraction is closely related to sample collection, processing, transport, and storage. Any error in these steps may lead to inaccurate results.
ZJ26F0636401
危险品化学品经营许可证（带存储）