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DyeEx 2.0 Spin Kit

用于去除1–24个测序反应体系中未结合的染料终止剂

  • 快速的操作流程只需两步短暂离心
  • 即用型预水化凝胶过滤材料
  • 高效去除任何染料终止剂

DyeEx Kits提供离心柱或96孔板规格,应用凝胶过滤技术从测序反应体系中去除染料终止剂。将测序反应体系上载到预水化凝胶过滤材料中,短暂离心后,反应体系即可用于毛细管测序仪测序。未结合的染料终止剂则会滞留在凝胶基质中。

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Cat No./ID: 63204
DyeEx 2.0 Spin Kit (50)
50 DyeEx Spin Columns, Collection Tubes (2 ml)
Cat No./ID: 63206
DyeEx 2.0 Spin Kit (250)
250 DyeEx Spin Columns, Collection Tubes (2 ml)

DyeEx 2.0 Spin Kit适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


Product Details

9
DyeEx separation principle.
The DyeEx procedure uses gel filtration to quickly and efficiently remove unincorporated terminators from sequencing reactions.
9
Superior sequencing results.
[A] Gel images and [B] sequence profiles of QIAprep purified plasmids sequenced using the ABI PRISM BigDye Terminator Cycle Sequencing Kit and ABI PRISM 377 DNA Sequencer. Sequencing reactions were purified using the DyeEx 96 Kit or precipitated with ethanol, as recommended by the supplier of the sequencing kit. The plasmids sequenced were plasmids from a genomic library (ethanol precipitation, gel image) or were pUC21 (DyeEx 96 Kit and ethanol precipitation, sequence profiles). Dye terminators present in ethanol-precipitated sequencing reactions cause dye blobs (indicated by arrows) resulting in unreadable sequences.
9
Long-read sequence.
Sequence of a 4.7 kb plasmid sequenced using the ABI PRISM BigDye Terminator Sequencing Kit and purified using the DyeEx 96 Kit.
9
DyeEx 2.0 Spin procedure.
A quick centrifugation step removes storage buffer from the columns, the sequencing samples are loaded, and a second centrifugation step removes unincorporated dye terminators.
Performance

DyeEx Kits可去除10–20 µl测序反应体系中任意类型的染料终止剂,包括BigDye、dRhodamine染料、Rhodamine、DYEnamic ET和WellRED终止剂。片段回收后,测序反应可在任意类型的毛细管测序仪上完成。

高品质测序

DyeEx Kits经优化,用于快速简便的去除染料终止剂,确保高品质的测序结果。与之相反,通过乙醇沉淀进行测序反应体系的回收非常耗时(参见"Comparison of DyeEx procedure and ethanol precipitation")并且效率较低。无法高效去除染料终止剂将会导致测序数据上出现染斑,使得延伸序列不可读。使用DyeEx Kits进行反应体系的回收,可以确保上载到测序设备的反应体系不含染料终止剂。无染斑干扰,可轻松读通序列(参见"Superior sequencing results")。信号强度高可确保较长的序列读取长度(参见"Long-read sequence")。

DyeEx流程与乙醇沉淀的比较
操作流程  DyeEx 2.0 Spin Kit 乙醇沉淀
所需时间*
(12 个样本,离心规格)
10分钟 >45分钟
所需时间*
(用于96个样本的微孔板规格)
N.A. >60分钟
处理 即用型离心规格 多步移液操作
测序品质 ++  +
N.A.: 不适用
* 不包括凝胶装载前干燥样本所需时间(如适用).
使用标准的实验方案.
Principle
DyeEx染料终止剂去除系统通过简单的离心柱流程,配合该流程中的凝胶过滤,快速高效的去除测序反应体系中未结合的染料终止剂。为了避免未结合的染料终止剂干扰测序结果的分析,因此去除染料终止剂非常重要。DyeEx凝胶过滤材料由带有均匀孔隙的球体组成,根据分子量对染料分子进行分离。当测序反应混合物上样到DyeEx柱的时候,染料终止剂扩散到孔隙内,并滞留在凝胶过滤材料中,而标记的DNA片段从球体间隙通过,由流出液中回收得到(参见"DyeEx separation principle")。
Procedure
DyeEx Kits可快速除去染料终止剂,因为其流程非常简单(参见"DyeEx 2.0 Spin procedure")。通过快速离心除去柱子中残留的缓冲液,然后上载测序样本,接着通过第二步离心除去未结合的染料终止剂。样本此时即可直接上载到毛细测序仪上,或经干燥,再溶解后上样到测序凝胶。
Applications

回收后,测序反应可在任意毛细管测序仪上完成。

Specifications

Features
Specifications
结合能力 10–20 µl
洗脱体积 10–20 µl
用于DyeEx Kits: Terminator removal, ABI PRISM 377, 373, 310, 3100, or 3700, MegaBACE 1000, or CEQ 2000 sequencers
规格 Tube/96-well plate
处理 Manual
去除<10mers、17–40mers的染料终止子蛋白 Dye terminator
样本类型:应用 Sequencing reactions
技术 Gel filtration

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