Omniscript RT Kit

For reverse transcription of 50 ng to 2 μg RNA per reaction for end-point PCR

  • High cDNA yields due to high-affinity enzyme
  • Sensitive detection of as few as 10 copies of template
  • No additional RNase H digestion step required
  • Fast and easy procedure with no tedious pipetting steps
The Omniscript Reverse Transcription (RT) Kit is specially designed for reverse transcription with any amount of RNA from 50 ng to 2 μg per reaction. A high affinity for RNA allows Omniscript Reverse Transcriptase to provide superior performance compared with other reverse transcriptases, delivering higher sensitivity in RT-PCR, even with low-copy numbers. The Omniscript RT Kit includes Omniscript Reverse Transcriptase, dNTPs and an optimized reaction buffer. Simply add primers for fast and easy cDNA synthesis.
产品 货号 目录价:
Omniscript RT Kit (50)
For 50 reverse-transcription reactions: 200 units Omniscript Reverse Transcriptase, 150 µl 10x Buffer RT, 100 µl dNTP Mix (contains 5 mM each dNTP), 1.1 ml RNase-free water
205111
询价
Omniscript RT Kit (200)
For 200 reverse-transcription reactions: 800 units Omniscript Reverse Transcriptase, 4 x 150 µl 10x Buffer RT, 4 x 100 µl dNTP Mix (contains 5 mM each dNTP), 4 x 1.1 ml RNase-free water
205113
询价

Omniscript RT 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。


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全长的RT-PCR产物— A。|全长的RT-PCR产物— B。|Omniscript Reverse Transcriptase实验流程。|多种逆转录酶的对比。|对10个以上拷贝进行灵敏的RT-PCR。|
采用图示的RT反应温度,对从玉米叶中提取的总RNA进行RT-PCR。扩增1.2 kb的富含GC的玉米gl2转录本片段。采用标准的Omniscript RT实验方案,按说明在标准(37°C)和较高的反应温度下进行RT。|采用图示的RT反应温度,对从玉米叶中提取的总RNA进行RT-PCR。扩增1.2 kb的富含GC的玉米gl2转录本片段。采用标准的MMLV RNase H- RT实验方案(Supplier L),按说明在标准(42°C)及其他反应温度下进行RT。该标准实验方案包括必需的初步变性步骤以及在RT结束后推荐进行的RNase H酶酶切步骤。|Omniscript RT Kit包括dNTPs和优化的反应溶液以及对RNA高亲和性的Omniscript Reverse Transcriptase,能够通读高GC含量和二级结构复杂的序列。由于这种预优化,无需繁琐的移液和预孵育步骤,也无需额外的RNase H消化步骤。|使用从HeLa细胞中提取的图示量的总RNA,依照供应商提供的说明书使用不同的逆转录酶进行逆转录。1/20的逆转录反应使用QIAGEN Taq DNA Polymerase进行25个循环的PCR扩增反应。扩增1.7 kb的β-actin片段。Omniscrip:Omniscript Reverse Transcriptase(QIAGEN);MMLV RNase H–:来自Moloney小鼠白血病病毒的RNase H–逆转录酶(供应商L);MMLV:野生型MMLV逆转录酶(供应商L);AMV RNase H–:来自禽成髓细胞瘤病毒的RNase H–逆转录酶(供应商L);AMV:野生型AMV逆转录酶(供应商P)。|从10到104拷贝的起始模板中(在200 ng总RNA中)逆转录一个700 nt的体外转录子,随后进行PCR。每个逆转录反应的1/4(相当于2.5到2500拷贝的 cDNA)用于PCR扩增一个330 bp的片段。|
性能

The high affinity of Omniscript Reverse Transcriptase (RT) results in highly specific and sensitive RT-PCR, even with low-copy transcripts (see figure "Sensitive RT-PCR of ≥10 copies"), and the ability to read through even complex RNA secondary structures without adjusting temperature or reaction conditions (see figure "Comparison of various reverse transcriptases").

Regions of RNA with high GC content can cause other reverse transcriptases to stop, dissociate from the RNA template, or skip over looped-out regions of RNA (see figure "Full-length RT-PCR products — B"). These difficult templates, however, prove no problem for QIAGEN reverse transcriptases (see figure "Full-length RT-PCR products — A"). With no need for optimization, the Omniscript RT Kit makes reverse transcription virtually trouble-free.

原理

Omniscript Reverse Transcriptase (RT) has a high affinity for RNA, which enables efficient and sensitive reverse transcription of any template, leading to high yields of cDNA. It is provided ready-to-use with dNTPs and in an optimized reaction buffer that, together with the high affinity of Omniscript RT for RNA, enables read-through of templates with high GC content or complex secondary structure. Please note that the primer mix is not provided.

Omniscript RT is specially designed for all reverse transcription with any amount of RNA from 50 ng to 2 µg per reaction. Omniscript RT is also usually the enzyme of choice with viral RNA due to the presence of carrier RNA in most viral RNA preparations. In comparative experiments, Omniscript RT consistently outperforms other reverse transcriptases over a wide range of starting RNA amounts.

Lot-to-lot reproducibility of Omniscript Kits is ensured by rigorous quality control at QIAGEN. The optimized Buffer RT, dNTPs, and water included in all Omniscript RT Kits are guaranteed RNase-free, and each lot of Omniscript RT is thoroughly tested for RT-PCR reproducibility.

操作流程
With the optimized Omniscript Reverse Transcriptase reaction buffer, tedious pipetting and pre-incubation steps are eliminated, and no additional RNase H digestion step is needed (see flowchart "Omniscript Reverse Transcriptase Procedure").
应用

Omniscript Reverse Transcriptase is suitable for use in the following applications:

  • cDNA synthesis for end-point PCR
  • Synthesis of double-stranded cDNA for cloning
  • RACE (Rapid Amplification of cDNA Ends)
  • Linear RNA amplification
  • Exponential RNA amplification
  • SAGE (Serial Analysis of Gene Expression)
  • Labeling for microarrays
  • Analysis of transcription start site by primer extension

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图片
Full-Length RT-PCR Products.
全长的RT-PCR产物— A。
采用图示的RT反应温度,对从玉米叶中提取的总RNA进行RT-PCR。扩增1.2 kb的富含GC的玉米gl2转录本片段。采用标准的Omniscript RT实验方案,按说明在标准(37°C)和较高的反应温度下进行RT。
Full-Length RT-PCR Products.
全长的RT-PCR产物— B。
采用图示的RT反应温度,对从玉米叶中提取的总RNA进行RT-PCR。扩增1.2 kb的富含GC的玉米gl2转录本片段。采用标准的MMLV RNase H- RT实验方案(Supplier L),按说明在标准(42°C)及其他反应温度下进行RT。该标准实验方案包括必需的初步变性步骤以及在RT结束后推荐进行的RNase H酶酶切步骤。
Omniscript RT procedure
Omniscript Reverse Transcriptase实验流程。
Omniscript RT Kit包括dNTPs和优化的反应溶液以及对RNA高亲和性的Omniscript Reverse Transcriptase,能够通读高GC含量和二级结构复杂的序列。由于这种预优化,无需繁琐的移液和预孵育步骤,也无需额外的RNase H消化步骤。
Superior sensitivity and dynamic range of Omniscript RT
多种逆转录酶的对比。
使用从HeLa细胞中提取的图示量的总RNA,依照供应商提供的说明书使用不同的逆转录酶进行逆转录。1/20的逆转录反应使用QIAGEN Taq DNA Polymerase进行25个循环的PCR扩增反应。扩增1.7 kb的β-actin片段。Omniscrip:Omniscript Reverse Transcriptase(QIAGEN);MMLV RNase H–:来自Moloney小鼠白血病病毒的RNase H–逆转录酶(供应商L);MMLV:野生型MMLV逆转录酶(供应商L);AMV RNase H–:来自禽成髓细胞瘤病毒的RNase H–逆转录酶(供应商L);AMV:野生型AMV逆转录酶(供应商P)。
Sensitive RT-PCR of ≥10 Copies
对10个以上拷贝进行灵敏的RT-PCR。
从10到104拷贝的起始模板中(在200 ng总RNA中)逆转录一个700 nt的体外转录子,随后进行PCR。每个逆转录反应的1/4(相当于2.5到2500拷贝的 cDNA)用于PCR扩增一个330 bp的片段。