手机验证
4℃
有效期1年
Hieff NGSTM DNA Selection Beads(Superior AMPure XP Alternative)
大量
翌圣生物科技(上海)股份有限公司
N/A
1ml
产品信息
产品名称 |
产品编号 |
规格 |
价格(元) |
Hieff NGS® DNA Selection Beads DNA分选磁珠 |
12601ES03 |
1 mL |
296.00 |
12601ES08 |
5 mL |
986.00 |
|
12601ES56 |
60 mL |
6286.00 |
|
12601ES75 |
450 mL |
26186.00 |
产品描述
Hieff NGS® DNA Selection Beads基于SPRI (Solid Phase Reverse Immobilization)原理,配合精心优化过的缓冲体系,可用于二代测序文库构建过程中的DNA片段分选、纯化。本产品可适用于各品牌的DNA、RNA建库试剂盒,和目前广泛使用的AMPure XP Beads使用方式相同,片段回收效率和文库大小分布均与AMPure XP Beads高度吻合。
运输与保存方法
冰袋运输。2-8℃保存,效期一年。避免冷冻!
注意事项
1)为了您的安全和健康,请穿实验服并戴一次性手套操作。
2)磁珠使用前须在室温平衡至少30 min。
3)80%乙醇需现用现配,否则将影响回收效率。
4)进行长度分选时,初始样品体积需≥100μL,不足时请用超纯水补齐。样品体积太小,将导致移液误差增大,进而影响分选的准确性。
5) 本产品仅用作科研用途!
使用方法
1. 准备工作
将磁珠由冰箱中取出,室温平衡至少30 min。配制80%乙醇。
2. 长度分选(双轮法)
长度分选操作流程如图1所示,具体操作如下。
图1双轮分选操作流程
1)请涡旋振荡或充分颠倒磁珠以保证混匀。
2)根据要求,参考表1向DNA溶液中加入第一轮分选磁珠,涡旋混匀或移液器吹打10次混匀。
3)室温孵育5 min。
4)将离心管短暂离心并置于磁力架中,待溶液澄清后(约5 min),小心转移上清到干净的离心管中。
注意:转移上清时,请残留2 μL液体于管底,切勿全部吸出上清,避免吸到磁珠并影响分选效果。
举例:当初始体积为100 μL,第一轮使用0.8×比例,即加入80 μL磁珠,推荐吸出178 μL的上清。
5)参考表1向上清中加入第二轮分选磁珠。
6)涡旋混匀或移液器吹打10次混匀,室温静置5 min。
7)将离心管短暂离心并置于磁力架中,待溶液澄清后(约5 min),小心移除上清。
8)保持离心管始终处于磁力架中,加入200 μL新鲜配制的80%乙醇漂洗磁珠,室温孵育30 s,小心移除上清。
9)重复步骤8。
10)保持离心管始终处于磁力架中,开盖干燥磁珠至刚刚出现龟裂(约5 min)。
注意:切记磁珠不要干燥时间太久,磁珠干燥过度将影响纯化效果。
11)将离心管从磁力架中取出,加入适量ddH2O(≥20 μL),涡旋振荡或使用移液器轻轻吹打充分混匀,室温孵育5 min。
12)将离心管短暂离心并置于磁力架中分离磁珠和液体。待溶液澄清后(约5 min),小心吸取上清至干净的管中,即完成分选。
3. DNA片段分选参考条件
通过超声法将小牛胸腺DNA进行片段化,制备100-1000 bp的Smear片段,根据表1进行双轮分选。结果使用Agilent 2100 Bioanalyzer进行分析(图2)。
表1磁珠文库分选推荐比例
DNA片段大小 | 250-350 bp | 320-420 bp | 450-550 bp | 550-700 bp | 700-900 bp | 800-1000 bp |
第一轮体积比(Beads:DNA) | 0.80× | 0.70× | 0.60× | 0.55× | 0.50× | 0.45× |
第二轮体积比(Beads:DNA) | 0.20× | 0.20× | 0.20× | 0.15× | 0.15× | 0.15× |
注:表中“×”表示样品DNA体积。如文库插入片段长度为250 bp,样品DNA体积为100mL,则第一轮分选磁珠使用体积为0.80×100 mL=80 mL;第二轮分选磁珠使用体积为0.20×100 mL=20 mL。
图2 Agilent 2100 high sensitivity DNA chip electopherogram
Smear fragments溶于ddH2O,使用0.80×/0.20×至0.45×/0.15×磁珠进行片段分选
HB220528
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