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101Bio//Bio

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货号:Bio
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品牌:101Bio
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商品描述
2x Es Taq Master Mix (with dye)

Name

2x Es Taq Master Mix (with dye)

Cat. #

W0690-1
$ 39.00
(1 mL, 100 rxn, 20 ul/rxn)
W0690-5
$ 99.00
(5 mL, 500 rxn, 20 ul/rxn)
How to pay with

Application

  • Routine PCR
  • T/A cloning
This product is for research use only.

Specifications

Polymerase:Es Taq
Hot Start:N/A
Fidelity:High fidelity
GC-Rich PCR Performance:High
Amplification Range:6 kb
Exonuclease Activity:5´→3´
Reaction Speed:1 kb / 30 seconds
Reaction Format:Master Mix
Product Overhang:3"-A

Description and features

This Master Mix contains Es Taq DNA Polymerase, PCR buffer, Mg2+, dNTP, PCR stabilizer and PCR enhancer.The concentration is 2x.

Es Taq DNA Polymerase possesses 5´→3´ DNA polymerase and 5´→3´ exonuclease activity. The polymerase has the high amplification efficiency and low mismatching as Taq DNA polymerase and high fidelity of Pfu DNA polymerase. Es Taq Polymerase catalyzes the non-template directed addition of an adenine residue to the 3´-end of both strands of DNA molecules to make it suitable for T/A cloning.The amplification range of Es Taq is ~ 6 kb.

This unique Master Mix recipe makes the system very reliable.More than 98% of PCR reaction can get successful amplification during the first try.It also works well on complicate templates.

The Master Mix contains dye, and can directly run electrophoresis after PCR reaction.

Shipping / Storage

Ship at 4℃. Store at -20℃ for up to 1 year and avoid freeze-thaw cycles. Stored at 4℃ for up to 3 months.

Quality control

This product is tested for no exogenous nuclease activity;no host DNA contamination tested (by PCR);able to amplify single copy gene from multiple genomes; and no significant enzyme activity decrease after storing at 2 ~ 8oC for 3 months.

Manual (protocol)

101Bio.com 2x Es Taq Master Mix

Components

ComponentsAmount
W0690-1W0690-5
2x Es Taq Master Mix1 mL1 mL x 5
RNase-Free Water1 mL5 mL

PCR reaction system

2x Es Taq Master Mix, convenient, save time and reliable

Note: The recommended primer concentration for PCR is between 0.1-1.0 µM of each primer. The use of higher concentrations of primers can have higher amplification effect. Low primer concentration will generally ensure cleaner product and lower background.

PCR reaction conditions

2x Es Taq Master Mix, convenient, save time and reliable

Note:

  • The recommended annealing temperature is about 5℃ below Tm of primers. If extra bands are observed, higher annealing temperatures should be considered. The absence of product indicates the need for a lower annealing temperature.
  • PCR extension time is depended on the size of target gene sequence.Es Taq DNA polymerase is approximately 1 kb DNA / 30 seconds.
  • The number of PCR cycles will basically depend on the downstream application of the PCR product.

PCR result examination

This Master Mix contains dye for electrophoresis.After PCR, directly load 5 µL of PCR product to agarose gel to run electrophoresis.No need to add loading buffer.

Exosome IsolationPurity > 95%
Protein Extraction1 min total protein, 40 min membrane protein
3D Cell Culture Gel30% < mkt="">
PCR Kits50% < mkt="">
Beta-Hexosaminidase Activity Colorimetric Assay Fast and sensitive, High-throughput
Endotoxin-Free Plasmid Kitsmaxi, midi and mini-prep
101bio外泌体提取试剂盒在获得极低白蛋白含量的外泌体中的应用外泌体(Exosomes)是大多数正常细胞和病变细胞都能够分泌的一种细胞外囊泡,粒子直径大约在30-150nm之间。外泌体广泛存在于血浆、乳液、尿液、唾液等体液中,并可携带母细胞中的多种蛋白、脂类、DNA、RNA等物质,参与细胞通讯、细胞迁移、血管新生和肿瘤细胞生长等过程。外泌体最早在19世纪80年代就有记载,当时人们只把它当做一种细胞废弃物。2013年诺贝尔生理学或医学奖被授予James E. Rothman等科学家,以表彰他们发现囊泡转运的调控机制。近年来,由于外泌体的特殊性质,研究人员对外泌体的研究热情激增,并发现其在诊断与治疗领域存在巨大潜力。在外泌体研究中,外泌体的分离提取是一直困扰广大研究人员的一大难题。外泌体领域资深科学家Clotilde Théry所建立的超速离心提取法一直被奉为外泌体提取的“金标准”。但超速离心法由于需要样本量大,耗时长,依赖特定仪器,提取量低,获得外泌体纯度低等缺点,大大降低了研究人员的研究效率。随着外泌体研究的不断发展,一些商品化试剂盒应运而生,这些试剂盒大多采用PEG沉淀原理,只需在半个小时左右的时间就可完成提取。但由于提取过程中PEG的加入,使获得的外泌体中含有PEG偶联物,严重影响后续实验的进行。另外,无论是超速离心法还是PEG沉淀试剂盒,所分离的外泌体中都含有大量的白蛋白,这也给后续的实验造成很大的干扰。针对外泌体提取过程所遇见的问题,美国101bio公司研发并生产了一款基于密度梯度分离法的外泌体分离试剂盒。该试剂盒相对于密度梯度离心省去了超速离心的步骤,而且还利用了外泌体特定极性和所带电荷等特性,比传统的密度梯度离心要简单很多,产量也高很多。最主要的是,通过对比101bio外泌体提取试剂盒所提取的外泌体白蛋白含量远远低于超速离心法和PEG沉淀法试剂盒所提取的外泌体,这样为研究人员后续实验扫清了一大障碍。另外,通过粒径分析,101bio外泌体提取试剂盒所提取的微粒直径在30-150nm之间的高达96.3%,说明该试剂盒所提取的外泌体几乎不包含其他类型微粒。