高通量测序产物纯化及片段筛选专用磁珠
Source:羧基磁珠Author:admin Addtime:2019/01/03 Click:
是一款专门为高通量测序而设计的磁珠试剂,同时满足对目的片段的纯化和目的片段筛选;既可以手工操作,也可以使用96孔或384孔板的形式完成。
产品优势:
1. 优异的缓冲体系,100bp以上扩增产物回收效率更高(100bp-10kb);
2. 有效去除dNTP、引物、引物二聚体、盐离子和其他杂质;
3. 磁珠磁含量更高,磁吸附时间更短;
4. 既适用于DNA纯化,也适用于RNA纯化;
5. 可配套自动化设备进行高通量产物纯化;
6. 手工操作和自动化工作站操作均可适用;
7. 无RNA酶的生产过程确保能应用于各种RNA的纯化操作。
产品应用:PCR产物纯化、目的片段筛选
操作过程:
一、纯化
1、添加磁珠和NGS文库混合,或者将PCR产物和磁珠混合
2、吸附已经和杂质分离的磁珠,并且吸出含杂质溶液
3、加入70%浓度的乙醇洗脱,然后吸附磁珠,重复该步骤
4、添加水,洗脱吸附在磁珠上的DNA,吸附磁珠,倒出纯化后的PCR产物
二、片段筛选
1、添加NGS吸附大片段
2、用磁铁吸附磁珠,使其从溶液中分离
3、将含有较小片段的上清液转移到干净的试管中
4、第二次加入磁珠,使其和需要的片段结合
5、用磁铁吸住磁珠,倒弃有杂质的上清
6、用80%乙醇洗脱磁珠
7、添加水进行洗脱,使用磁力架,吸附磁珠,并倒出纯化和筛选好的DNA片段
结果展示:
CleanNGS vs Competitor A(爱森 Single Cell Sequencing)
sample ID |
片段筛选磁珠(爱森) |
Competitor A |
avg size (bp) |
conc (ng/ul) |
avg size (bp) |
conc (ng/ul) |
1 |
696 |
26.7 |
691 |
25.9 |
2 |
580 |
27.1 |
587 |
25.3 |
3 |
569 |
27.6 |
567 |
25.4 |
4 |
607 |
28.3 |
603 |
27.1 |
爱森 vs Competitor A(Control for qPCR inhibition)
sample ID |
爱森 |
Competitor A |
NTC |
Ct (1x) |
18.00 |
21.23 |
29.47 |
19.56 |
21.39 |
28.70 |
20.06 |
20.82 |
28.62 |
|
Human genomic DNA sheared to 150, 200, 400 and 1000 bp fragments using the Covaris S2.
Fragments have been pooled and 10 µL of 1.5 – 1.8 ng/µL sheared genomic DNA has been purified using 1.8x ratio (18 µL) of the appropriate beads. Nuclease free water was used as a NTC. After binding the beads were washed twice using 80% ethanol and dried at RT for 5 minutes.
An on-bead qPCR was performed after adding 20 µL of SYBR master mix to the bead pellet containing the purified genomic DNA. |
更多规格:
包装规格 |
测试数量 |
货号 |
5mL |
277 |
AS316011 |
50mL |
2777 |
AS316011 |
500mL |
27777 |
AS316011 |
Number of reactions is based on a typical 10 µL PCR reaction volume.
Volume of CleanNGS to be used per reaction = 1.8x the sample volume.
www.ex-dna.com 磁珠法核酸提取试剂盒