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- 详细信息
- 文献和实验
- 技术资料
- 库存:
大量
- 英文名:
Moloney Murine Leukemia Virus Reverse Trancscriptase Recombinant
- 保质期:
1年
- 供应商:
上海沪震实业有限公司
- 保存条件:
-20°C
- 规格:
2500U/10000U/50000U
CATALOGUE NUMBER
ENZ-310
DESCRIPTION
MMLV (Moloney Murine Leukemia Virus) Reverse Transcriptase is a DNA polymerase that synthesizes a complementary DNA strands from single-stranded RNA, DNA, or an RNA-DNA hybrid as a template. This recombinant enzyme was purified from E.coli, which carried a modified MMLV-RT gene. Compared to AMV Reverse Transcriptase, this enzyme has a much weaker 5' - 3' ribonuclease H activity, which allows the syntesis of longer cDNAs (>7kb).SOURCE
Recombinant E. coli strain.PHYSICAL APPEARANCE
Sterile Filtered clear solution (200 U/µl).FORMULATION
50mM Tris-HCl, 0.1M NaCl, 0.1% Triton X-100, 2mM DTT, 0.1mM EDTA and 50% glycerol.STABILITY
Stable for 5 days at 10°C, for longer period of time store at -20°C.Please prevent freeze-thaw cycles.
UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 1nmol of deoxyribonucleotide into acid-insoluble forms in 10 minutes at 37oC, using poly(A)-oligo(dT)12-18 as the template-primer. Standard cDNA Synthesis Conditions50mM Tris-HCl (pH8.3), 75mM KCl, 3mM MgCl2, 10mM DTT, 1.0mM each dATP, dGTP, dCTP, and dTTP, 0.2 mg radom hexamer,1-5mg RNA, 200units M-MLV RT. The reaction volume was 20ml and the incubation was 45 min at 42oC.5X REACTION BUFFER
250mM Tris-HCl (pH8.3), 375mM KCl, 15mM MgCl2, and 50mM DTT.USAGE
Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.风险提示:丁香通仅作为第三方平台,为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全,合理判断,谨慎选购商品,商家和用户对交易行为负责。对于医疗器械类产品,请先查证核实企业经营资质和医疗器械产品注册证情况。
文献和实验合酶活性相互竞争RNA模板与DNA引物或cDNA延伸链间形成的杂合链,并降解RNA:DNA复合物中的RNA链。被RNaseH活性所降解的RNA模板不能再作为合成cDNA的有效底物,降低了cDNA合成的产量和长度。因此消除或大大降低逆转录酶的RNaseH活性将会大有裨益。SuperScriptⅡ逆转录酶,RNaseH- 的MMLV逆转录酶及ThermoScript逆转录酶,RNaseH- 的 AMV,比MMLV和AMV得到更多量和更多全长的cDNA(图3)。RT-PCR灵敏度会受cDNA合成量的影响
随机引物 适用于长的或具有发卡结构的 RNA。适用于 rRNA、mRNA、tRNA 等所有 RNA 的反转录反应。主要用于单一模板的 RT-PCR 反应。 Oligo dT 适用于具有 PolyA 尾巴的 RNA。(原核生物的 RNA、真核生物的 Oligo dT rRNA 和 tRNA 不具PolyA 尾 巴。)由于 Oligo dT 要结合到 PolyA 尾巴上,所以对 RNA 样品的质量要求较高,即使有少量降解也 会使全长 cDNA 合成量大大减少。 基因特异性引物 与模板序列互补
定量RT-PCR (Quantitative RT-PCR)
A = B(1+e) n A=amplified products, B=input templates, n=cycle number, and e=amplification efficiency. Factors affecting amplification efficiency in the RT-PCR process include the efficiency of reverse
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