 连接被互补 RNA 固定的相邻的单链 DNA
 鉴定 miRNAs 和 mRNAs,包括 SNPs  

SplintR 连接酶也称为 PBCV-1 DNA 连接酶或 Chorella 病毒 DNA 连接酶，它能够高效催化相邻的 DNA 核苷酸的连接，这个连接过程需要一条互补的 RNA 在两条 DNA 单链间起“夹板”或“桥梁”的固定作用。这种以前没有报道过的活性可能推动一些创新性 miRNAs 和 mRNAs,包括 SNPs 的分析方法。在二代测序和分子诊断等众多领域中，SplintR 是富集目的基因的理想选择。它具有稳定的生物活性，对 RNA 介导固定的 DNA 底物亲和性强（米氏常数 Km = 1 nM），能够在复杂的混合物中检测到亚纳摩尔级的特定 RNA。因此，在 RNA 检测技术中，SplintR 连接酶不失为最佳选择用酶。  

来自重组的 E. coli 菌株，其携带编码 PBCV-1 DNA 连接酶的基因。 

1X SplintR 连接酶反应缓冲液，25℃ 温育，最佳连接温度是 16℃。热失活：65℃ 温育 20分钟。  

SplintR 连接酶经过严格的质控检测，确保该产品具有最高的活性和纯度。详情请登陆 www.neb.com 或 www.neb-china.com。  

1 单位指 20 μl 反应体系中，1X SplintR 连接酶反应缓冲液，16℃ 条件下，15 分钟内能使 2 pmol（完成 50%）三重 FAM 标记的 DNA:RNA 杂交底物连接所需的酶量。单位活性检测条件请登陆 www.neb.com 或 www.neb-china.com。  

25,000 units/ml。 

关于该酶特性和产品应用的参考文献，请登陆 www.neb.com 或 www.neb-china.com。  

   Recombinant enzyme with no detectable endoglycosidase or other exoglycosidase contaminating

      activities

   Glycerol -free for optimal performance in HPLC and mass spectrometry analysis

   ≥95% purity, as determined by SDS-PAGE and intact ESI-MS

α1-2,4,6 Fucosidase O is a broad specificity exoglycosidase that catalyzes the hydrolysis of terminal

α1-2, α1-4 and α1-6 linked fucose residues from oligosaccharides. α1-2,4,6 Fucosidase O cleaves

α1-6 fucose residues more efficiently than other linkages.

Cloned from Omnitrophica bacterium and expressed in E. coli ⁽¹⁾.

1X GlycoBuffer 1.  Incubate at 37°C.

Heat Inactivation: 65°C for 10 min

10X GlycoBuffer 1 

1X GlycoBuffer 1
5 mM CaCl2
50 mM sodium acetate
(pH 5.5 @ 25°C)

Apparent: 49 kDa

One unit is defined as the amount of enzyme required to cleave > 95% of the fucose from 1 nmol of G0F

from human IgG [GlcNAcβ1-2Manα1-6(GlcNAcβ1-2Manα1-3)Manβ1-4GlcNAcβ1-4GlcNAc(Fucα1-6)-

AMAC], in 1 hour at 37°C in a total reaction volume of 10 µl. 

4°C

1.   Reactions may be scaled-up linearly to accommodate larger reaction volumes.
2.   The enzymatic unit definition is calculated using a 37°C reaction temperature, to accommodate for

      simultaneous glycosidase digestions and N-glycan sequencing protocols. However, the optimal

      reaction temperature for α1-2,4,6 Fucosidase O is 50°C. A 37°C reaction temperature results in

      ~35% lower activity.
3.   In order to achieve complete removal of core α1-6 fucose residues from glycans with instant labels,

      it may be necessary to use longer incubation times (18 hours).
4.   α1-2,4,6 Fucosidase O is only active on an intact antibody if it is trimmed to the trimannosyl core.

1.   Vainauskas, S., New England Biolabs, Inc. Unpublished observation
2.   Wong-Madden, S.T. and Landry, D. (1995). Glycobiology. 5, 19-28.