Fragments separation magnetic beads NGSbead | NGS magnetic beads kit | | the second generation sequencing magnetic beads Agencourt SPRIselect equivalent magnetic beads
| 【Numbering】BK2021081227 | 【CAS】 | ||
| 【Item No.】 | 【specification】 | ||
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NGSbead magnetic beads are designed for the selective or non-selective recovery of DNA when the second-generation sequencing database is built, as well as the purification and recovery of PCR products. The fragment sorting kit is suitable for rapid sorting and recycling of DNA fragment sizes in the construction of high-throughput sequencing libraries (NGS). NGSbead is directly dispersed in the binding buffer, adjusting the volume ratio of magnetic bead suspension and sample, and the left, right or bilateral size of DNA fragments can be selected. The purified fragments contained no contaminants such as nucleotides, primer dimers, joints, enzymes and salts. NGSbead provides an efficient purification system based on magnetic beads for the new generation of sequencing workflow. It is a large-scale and high-throughput fragment sorting kit specially developed for supporting automated workbench, which can be applied to almost all automated instruments, and can also be manually separated from a small number of samples.
■ Product performance
The purity of the eluted DNA was high, and the ratio of A260/A280 ranged from 1.7 to 1.9, and the ratio of A260/A230 was usually above 2.0. The recovery rate of magnetic beads is high, the recovery rate of 500bp is >90%, the recovery rate of 200bp is >80%, and the recovery rate of 100bp is >70%.
■ Application direction
Rapid sorting and recycling of DNA fragment size in the construction of high-throughput sequencing library (NGS);
You can select left, right, or bilateral sizes of DNA segments;
1.8 times the volume of suspension after the combination, washing, elution, can be used for PCR, enzyme reaction product purification.
■ Matters needing attention
1. Magnetic beads are stored in deionized water. Freezing, drying and centrifugation may affect the use effect of magnetic beads;
2. It is normal for magnetic beads to settle due to gravity. Before use, be sure to fully oscillate and mix the magnetic beads;
■ Use method
Reagent composition
NGSbead: Vortex 20 seconds before use, make it thoroughly mixed.
Wash buffer: 80% ethanol (used in situ, stored at 4℃ for no more than 7 days);
Elution buffer: 10 mM Tris-HCl (PH 8.0) or deionized water;
1. The first magnetic bead sorting (removing large segments of DNA that are not needed)
1. NGSbead beads were placed at room temperature for 30 min, and the vortex was mixed until the bead suspension was uniform in color. Refer to Table 1, add an appropriate amount of magnetic bead solution into the reaction solution, blow at least 10 times to mix the mixture solution, and leave it at room temperature for 5 min.
Table 1 Ratio of magnetic beads suspension to sample volume (the amount of magnetic beads in the table is the relative residual sample volume)
2. Place the 96-well plate on the magnetic rack to separate the magnetic beads, and let stand for at least 5 minutes until the supernatant is completely clarified.
3. Transfer the supernatant to the new 96-well plate and discard the magnetic beads with large fragments of DNA. (Note: Do not discard the supernatant)
2. The second magnetic bead sorting (removing small fragments of DNA that are not needed)
1. Vortex resuspended and mixed NGSbead magnetic beads to uniform color, magnetic beads solution was added to the above supernatant (refer to Table 1), blowing at least 10 times to mix, standing at room temperature for 5 min.
2. Place the 96-well plate on the magnetic rack to separate the magnetic beads, and let stand for at least 5 minutes until the supernatant is completely clarified.
3. Carefully remove supernatants that contain unnecessary DNA. (Note: Do not discard magnetic beads
Three, ethanol cleaning
1. In the ethanol cleaning step, it is necessary to put the 96-well plate on the magnetic rack, and do not touch the magnetic beads during the cleaning process. Add 200 μL of 80% ethanol and let stand at room temperature for at least 30 s. Discard the supernatant. Repeat this step once.
Four, des
1. Place the 96-well plate on a magnetic rack and leave it at room temperature for 5 min to volatilize the residual ethanol. (Note: dry to the bottom of the tube without obvious droplets; Do not over-dry magnetic beads to prevent low DNA recovery efficiency).
2. Remove the 96-well plate from the magnetic rack, add 10-50 μL Elution buffer to re-suspend blowing or oscillate mixing magnetic beads, and place at room temperature for 2-5 min. (Elution buffer is better after preheating at 65-70℃)
3. Place the 96-well plate on the magnetic rack. After solution is cleared, transfer the supernatant to a new 96-well plate.
Note: 1. The procedure provided is the process of removing small fragments and large fragments and obtaining fragments in the middle range. If the purpose of the experiment is to remove the small fragment and recover the large fragment or remove the large fragment and recover the small fragment, the operation only needs to select the appropriate magnetic bead solution proportion. 2. The kit magnetic bead solution can be used for the recovery of 300 bp~800 bp fragment. 3. Because the size of the recovered fragment is related to the proportion of the magnetic bead solution and the sample, the accuracy of the sample addition is very important, and the sample volume should not be too small as far as possible to reduce the sampling error, generally in the range of 50 μL~150 μL.
Message
■ Product performance
The purity of the eluted DNA was high, and the ratio of A260/A280 ranged from 1.7 to 1.9, and the ratio of A260/A230 was usually above 2.0. The recovery rate of magnetic beads is high, the recovery rate of 500bp is >90%, the recovery rate of 200bp is >80%, and the recovery rate of 100bp is >70%.
■ Application direction
Rapid sorting and recycling of DNA fragment size in the construction of high-throughput sequencing library (NGS);
You can select left, right, or bilateral sizes of DNA segments;
1.8 times the volume of suspension after the combination, washing, elution, can be used for PCR, enzyme reaction product purification.
■ Matters needing attention
1. Magnetic beads are stored in deionized water. Freezing, drying and centrifugation may affect the use effect of magnetic beads;
2. It is normal for magnetic beads to settle due to gravity. Before use, be sure to fully oscillate and mix the magnetic beads;
■ Use method
Reagent composition
NGSbead: Vortex 20 seconds before use, make it thoroughly mixed.
Wash buffer: 80% ethanol (used in situ, stored at 4℃ for no more than 7 days);
Elution buffer: 10 mM Tris-HCl (PH 8.0) or deionized water;
1. The first magnetic bead sorting (removing large segments of DNA that are not needed)
1. NGSbead beads were placed at room temperature for 30 min, and the vortex was mixed until the bead suspension was uniform in color. Refer to Table 1, add an appropriate amount of magnetic bead solution into the reaction solution, blow at least 10 times to mix the mixture solution, and leave it at room temperature for 5 min.
Table 1 Ratio of magnetic beads suspension to sample volume (the amount of magnetic beads in the table is the relative residual sample volume)
2. Place the 96-well plate on the magnetic rack to separate the magnetic beads, and let stand for at least 5 minutes until the supernatant is completely clarified.
3. Transfer the supernatant to the new 96-well plate and discard the magnetic beads with large fragments of DNA. (Note: Do not discard the supernatant)
2. The second magnetic bead sorting (removing small fragments of DNA that are not needed)
1. Vortex resuspended and mixed NGSbead magnetic beads to uniform color, magnetic beads solution was added to the above supernatant (refer to Table 1), blowing at least 10 times to mix, standing at room temperature for 5 min.
2. Place the 96-well plate on the magnetic rack to separate the magnetic beads, and let stand for at least 5 minutes until the supernatant is completely clarified.
3. Carefully remove supernatants that contain unnecessary DNA. (Note: Do not discard magnetic beads
Three, ethanol cleaning
1. In the ethanol cleaning step, it is necessary to put the 96-well plate on the magnetic rack, and do not touch the magnetic beads during the cleaning process. Add 200 μL of 80% ethanol and let stand at room temperature for at least 30 s. Discard the supernatant. Repeat this step once.
Four, des
1. Place the 96-well plate on a magnetic rack and leave it at room temperature for 5 min to volatilize the residual ethanol. (Note: dry to the bottom of the tube without obvious droplets; Do not over-dry magnetic beads to prevent low DNA recovery efficiency).
2. Remove the 96-well plate from the magnetic rack, add 10-50 μL Elution buffer to re-suspend blowing or oscillate mixing magnetic beads, and place at room temperature for 2-5 min. (Elution buffer is better after preheating at 65-70℃)
3. Place the 96-well plate on the magnetic rack. After solution is cleared, transfer the supernatant to a new 96-well plate.
Note: 1. The procedure provided is the process of removing small fragments and large fragments and obtaining fragments in the middle range. If the purpose of the experiment is to remove the small fragment and recover the large fragment or remove the large fragment and recover the small fragment, the operation only needs to select the appropriate magnetic bead solution proportion. 2. The kit magnetic bead solution can be used for the recovery of 300 bp~800 bp fragment. 3. Because the size of the recovered fragment is related to the proportion of the magnetic bead solution and the sample, the accuracy of the sample addition is very important, and the sample volume should not be too small as far as possible to reduce the sampling error, generally in the range of 50 μL~150 μL.
| Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance |
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| Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance.The website pictures are from the Internet. The pictures are for reference only. Please take the real object as the standard. In case of infringement, please contact us to delete them immediately. |
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