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DyeEx Kit

适用于去除 1–24 或 1–96 测序反应中未结合的染料终止物

S_1084_5_GEN_V2

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✓ 博学专业的产品和技术支持

✓ 快速可靠的(再)订购

DyeEx 96 Kit (4)

Cat. No. / ID:   63181

4 个 DyeEx 96 孔板;4 个采样孔板,48 孔
PLN 3,357.00
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Kit
DyeEx Kit
DyeEx 2.0 Kit
Preparations
4 x 96
24 x 96
DyeEx Kit 旨在用于分子生物学应用。这些产品不能用于疾病诊断、预防和治疗。

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的(再)订购

特点

  • 程序很快,只需两个很短的离心步骤
  • 即用型预先水化凝胶过滤材料
  • 有效去除任何染料终止物

产品详情

DyeEx Kit 提供了离心柱或 96 孔板,使用凝胶过滤技术去除测序反应(包括用于 Sanger 测序的反应)中的染料终止物。测序反应被加载到预先水化的凝胶过滤材料上。经过短暂的离心步骤后,即可将反应装入毛细管测序仪。未结合的染料终止物会保留在凝胶基质中。

绩效

DyeEx Kit 可清除 10–20µl 测序反应中任何类型的染料终止物,包括 BigDye、dRhodamine 染料、Rhodamine、DYEnamic ET 和 WellRED 终止物。纯化后,测序反应可在任何毛细管测序仪上进行分离。我们推荐使用优化方案的 DyeEx 96 Kit。

高质量测序

DyeEx Kit 经过优化,可快速方便地去除染料终止物,从而获得高质量的测序结果。相比之下,用乙醇沉淀法纯化测序反应非常耗时(见表“DyeEx 程序与乙醇沉淀法的比较”)且效率低下。如果不能有效去除染料终止物,往往会导致测序数据中出现染料斑点,使序列片段无法读取。使用 DyeEx Kit 进行测序反应纯化可确保加载到测序仪器上的反应不含染料终止物。由于没有染料斑点,整个序列都能轻松读取(见图 “卓越的测序结果”)。信号强度高,因此读数很长(见图 “长读数序列”)。

DyeEx 程序与乙醇沉淀法的比较

程序DyeEx 2.0 SpinDyeEx 96乙醇
沉淀法
所需时间*
(12 个样本,离心规格)
10 分钟不适用>45 分钟
所需时间*†
(微孔板格式适用于 96 个样本)
不适用18 分钟>60 分钟
处理即用型
离心规格
即用型多个
移液步骤
序列质量+++++

N.A.:不适用

* 不包括凝胶加载前干燥样本所需的时间(如适用)。
†使用标准方案。

查看图表

原理

DyeEx 程序使用凝胶过滤法快速有效地去除测序反应中未结合的终止物。去除染料终止物对于防止未结合的染料终止物干扰测序结果分析非常重要。DyeEx 凝胶过滤材料由具有均匀孔隙的球体组成,可根据分子量分离分子。当测序反应混合物进入 DyeEx 柱时,染料终止物会扩散到孔中,并留存在凝胶过滤材料中,而标记的 DNA 片段则会被排除,并在流川液中回收(见图 “DyeEx 分离原理”)。

查看图表

程序

使用 DyeEx Kit 可以快速去除染料终止物,因为操作步骤非常简单(见流程图 “DyeEx 2.0 Spin 程序” ”DyeEx 96 Kit 程序”)。快速离心步骤可去除离心柱或孔中存储的缓冲液,然后加载测序样本,再进行第二次离心步骤去除未结合的染料终止物。然后样本就可以直接加载到毛细管测序仪上,也可以烘干、重新溶解并加载到测序凝胶上。

查看图表

应用

DyeEx Kit 可清除 10–20µl 测序反应中任何类型的染料终止物,包括 BigDye、dRhodamine 染料、Rhodamine、DYEnamic ET 和 WellRED 终止物。纯化后,测序反应可在任何毛细管测序仪上进行分离。信号强度高,因此读数很长。

DyeEx 特点

特点 DyeEx 2.0 Spin Kit 和 DyeEx 96 Kit
结合能力 10–20 µl
洗脱体积 10–20 µl
对于 DyeEx Kit: 终止物去除,ABI PRISM 377、373、310、3100 或 3700、MegaBACE 1000 或 CEQ 2000 测序仪
格式 试管 (DyeEx 2.0 Spin Kit)、96 孔板 (DyeEx 96 Kit)
处理 手动
去除 <10mer 17–40mer 染料终止物蛋白质 染料终止物
样本类型 测序反应
技术 凝胶过滤

DyeEx Kit 旨在用于分子生物学应用。这些产品不能用于疾病诊断、预防和治疗。

辅助数据和图表

资源

快速启动实验方案 (2)
DyeEx 96 Kit (EN)
PDF (455KB)
试剂盒操作手册 (1)
(EN) - DyeEx Handbook
PDF (1019KB)
For DyeEx 2.0 Spin Kit and DyeEx 96 Kit 
安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
Safety Data Sheets (1)
Certificates of Analysis (1)

Publications

Transcriptional regulatory network analysis of developing human erythroid progenitors reveals patterns of coregulation and potential transcriptional regulators.
Keller MA; Addya S; Vadigepalli R; Banini B; Delgrosso K; Huang H; Surrey S;
Physiol Genomics; 2006; 28 (1):114-28 2006 Aug 29 PMID:16940433
Isolation and identification of Rickettsia massiliae from Rhipicephalus sanguineus ticks collected in Arizona.
Eremeeva ME; Bosserman EA; Demma LJ; Zambrano ML; Blau DM; Dasch GA;
Appl Environ Microbiol; 2006; 72 (8):5569-77 2006 Aug PMID:16885311
Mammalian monogamy is not controlled by a single gene.
Fink S; Excoffier L; Heckel G;
Proc Natl Acad Sci U S A; 2006; 103 (29):10956-60 2006 Jul 10 PMID:16832060
High throughput analysis of TCR-beta rearrangement and gene expression in single T cells.
Zhou D; Srivastava R; Grummel V; Cepok S; Hartung HP; Hemmer B;
Lab Invest; 2006; 86 (3):314-21 2006 Mar PMID:16446708
Prevalence, risk factor analysis, and follow-up of infections caused by three feline hemoplasma species in cats in Switzerland.
Willi B; Boretti FS; Baumgartner C; Tasker S; Wenger B; Cattori V; Meli ML; Reusch CE; Lutz H; Hofmann-Lehmann R;
J Clin Microbiol; 2006; 44 (3):961-9 2006 Mar PMID:16517884

FAQ

Can the DyeEx 96 kit be used to clean sequencing reactions to be run on ABI 3730?

Yes, DyeEX 96 is also compatible with ABI 3730. When using DyeEx96 with ABI 3730, please follow the  "Modified protocol" listed in the DyeEX Handbook. In addition, if using the waterloading protocol on ABI 3730 sequencer,  the eluate can be loaded directly onto the sequencer without drying down the sample.

FAQ ID -3075
Can DyeEx be used to clean up labeled DNA other than sequencing templates?

DyeEx Kits have been developed for cleanup of dye-terminator sequencing reactions. However, they can also be used for removal of unincorporated nucleotides from radioactively labeled DNA fragments using the protocols in the DyeEx Handbook. The removal of nucleotides depends on the sample volume. Typically, more than 99% of nucleotides are removed from sample volumes ≤50 µl. See Figure 9 in the DyeEx Handbook for the effect of sample volume on recovery.

(We have had feedback from researchers who have reported excellent results using DyeEx for clean up of probes used for in situ hybridization).

FAQ ID -137
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