本试剂盒适用于从干血斑或血卡样本中提取高纯度的基因组DNA，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，得到的DNA产物可直接用于PCR、荧光定量PCR、芯片分析、一代、二代测序等相关实验。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 | | M1522695A | 蛋白酶K | 液体 | 1.1 mL | 2-8℃. | | M1522695B | 磁珠MBVB | 液体 | 1.1 mL | 2-8℃. | | M1522695C | 裂解液AL | 液体 | 30 mL | RT. | | M1522695D | 裂解结合液BLN | 液体 | 35 mL | RT. | | M1522695E | 洗涤液DW1 | 液体 | 40 mL | RT. | | M1522695F | 洗涤液DW2 | 液体 | 77 mL | RT. | | M1522695G | 洗脱液DE | 液体 | 5 mL | RT. |
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储存条件
M1522695A, M1522695B：2-8℃保存，12个月。
M1522695C, M1522695D, M1522695E, M1522695F, M1522695G：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：干血斑/血卡。
- 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
3. 试剂准备
4. 操作步骤
磁分离操作建议（步骤4.6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 用打孔器在FTA Card或血片上打3~5个3 mm直径的血片，如无打孔器，需将血片剪碎。转移至1.5 mL或2.0 mL离心管中。
4.2 加入400 μL裂解液AL、20 μL蛋白酶K，高速涡旋混匀30秒，检查确保血片浸没在溶液中。75℃振荡温浴45分钟，若恒温仪无振荡功能，可在温浴期间每15分钟取出涡旋1次，每次30秒。
4.3 待温浴后的溶液降至室温后，瞬时离心收集管盖液滴至管中。
4.4 将所有溶液转移至新的离心管中，避免吸到沉淀。
4.5 加入600 μL裂解结合液BLN、20 μL磁珠MBVB，振荡或涡旋5~7分钟使磁珠吸附DNA。
4.6 转移至磁力架上进行磁分离至溶液透亮，吸弃溶液。
4.7 加入700 μL洗涤液DW1，涡旋10秒打散磁珠后转至磁力架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秒打散磁珠后转至磁力架进行磁分离至溶液澄清，吸弃溶液。
4.9 再次加入700 μL洗涤液DW2，涡旋10秒打散磁珠后转至磁力架进行磁分离至溶液澄清，吸弃溶液。
4.10 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。
4.11 加入30~50 μL洗脱液DE，高速涡旋2~3分钟打散磁珠。
4.12 60℃温浴5分钟，然后高速涡旋30秒。
4.13 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。
4.14 把基因组DNA溶液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 is designed for the extraction of high‑purity genomic DNA from dried blood spots (DBS) or blood card samples. The method employs superparamagnetic bead purification technology, eliminating the need for toxic phenol‑chloroform extraction. The procedure is simple and convenient, and the resulting DNA can be used directly in PCR, real‑time PCR, microarray analysis, first‑generation and next‑generation sequencing, and related experiments. The purified DNA is intended for research use only.
| Product Code | Component | Appearance | 48T | Storage | | M1522695A | Proteinase K | Liquid | 1.1 mL | 2–8 °C | | M1522695B | Magnetic Beads MBVB | Liquid | 1.1 mL | 2–8 °C | | M1522695C | Lysis Buffer AL | Liquid | 30 mL | RT | | M1522695D | Lysis/Binding Buffer BLN | Liquid | 35 mL | RT | | M1522695E | Wash Buffer DW1 | Liquid | 40 mL | RT | | M1522695F | Wash Buffer DW2 | Liquid | 77 mL | RT | | M1522695G | Elution Buffer DE | Liquid | 5 mL | RT |
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Storage Conditions
M1522695A, M1522695B: Store at 2–8 °C for 12 months.
M1522695C, M1522695D, M1522695E, M1522695F, M1522695G: 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 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: Dried blood spots / blood cards.
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 mL sterile, nuclease‑free centrifuge tubes
Various pipettes and tips (20 µL, 200 µL, 1000 µL)
3. Reagent Preparation
Check solutions for precipitates and ensure magnetic beads can be resuspended.
4. Procedure
Recommended magnetic separation procedure (starting from step 4.6): 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 Use a hole punch to take 3–5 discs of 3 mm diameter from an FTA card or blood card. If a punch is not available, cut the blood spot into small pieces. Transfer the discs/pieces into a 1.5 mL or 2.0 mL centrifuge tube.
4.2 Add 400 µL of Lysis Buffer AL and 20 µL of Proteinase K. Vortex vigorously for 30 seconds to mix, ensuring the blood discs are submerged. Incubate at 75 °C with shaking for 45 minutes. If the incubator does not have a shaking function, remove the tube every 15 minutes and vortex for 30 seconds.
4.3 After incubation, cool the solution to room temperature. Briefly centrifuge to collect droplets from the cap.
4.4 Transfer the entire solution to a new centrifuge tube, avoiding any precipitate.
4.5 Add 600 µL of Lysis/Binding Buffer BLN and 20 µL of Magnetic Beads MBVB. Shake or vortex for 5–7 minutes to allow the beads to bind DNA.
4.6 Place the tube on a magnetic stand for magnetic separation until the solution is clear. Aspirate and discard the supernatant.
4.7 Add 700 µL of Wash Buffer DW1. Vortex for 10 seconds to resuspend the beads. Return to the magnetic stand and separate until the solution is clear. Aspirate and discard the supernatant.
4.8 Add 700 µL of Wash Buffer DW2. Vortex for 10 seconds to resuspend the beads. Separate magnetically until clear, then aspirate and discard the supernatant.
4.9 Add another 700 µL of Wash Buffer DW2. Vortex for 10 seconds to resuspend the beads. Separate magnetically until clear, then aspirate and discard the supernatant.
4.10 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.
4.11 Add 30–50 µL of Elution Buffer DE. Vortex vigorously for 2–3 minutes to fully resuspend the beads.
4.12 Incubate at 60 °C for 5 minutes, then vortex vigorously for 30 seconds.
4.13 Briefly centrifuge to collect droplets from the cap. Return the tube to the magnetic stand for magnetic separation.
4.14 Transfer the genomic DNA solution 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.
ZJ26F0636698
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