추천 제품
사용
sufficient for 150 reactions
sufficient for 50 reactions
Quality Level
특징
hotstart
제조업체/상표
Roche
포장
pkg of 150 reactions (04655885001)
pkg of 50 reactions (04655877001)
기술
RT-PCR: suitable
RT-qPCR: suitable
입력
purified RNA
검출 방법
probe-based
일반 설명
For hot start one-step RT-PCR, utilizing the enzyme mixture of Transcriptor Reverse Transcriptase, Protector RNase Inhibitor, and the Expand System, with an innovative hot start buffer (patents pending).
One-step RT-PCR is a very sensitive technique for determining the presence or absence of RNA templates, or to quantify the levels of expression through qualitative, semi-quantitative, or quantitative analysis of RNA transcription levels (e.g., for virus-level quantification). This RT-PCR method also facilitates the amplification of rare messages for cloning. The one-step format is convenient, shows reduced reaction-to-reaction variability, and prevents contamination by minimizing hands-on steps.
The Transcriptor One-Step RT-PCR Kit offers the outstanding benefits of Transcriptor Reverse Transcriptase, as well as the advantages of a hot start one-step RT-PCR format.
The Transcriptor One-Step RT-PCR Kit offers the outstanding benefits of Transcriptor Reverse Transcriptase, as well as the advantages of a hot start one-step RT-PCR format.
The Transcriptor One-Step RT-PCR Kit provides all components required for one-step RT-PCR (except primers and template) in a convenient format. The 50-reaction pack size also includes 10 control reactions (control RNA and control primer mix).
The kit′s RT-PCR Enzyme Mix contains four different enzymes: Transcriptor Reverse Transcriptase ensures sensitive and robust reverse transcription with high yield; Protector RNase Inhibitor, fully active at elevated temperatures, provides maximum template protection during reverse transcription; and the Expand System – a blend consisting of Taq DNA Polymerase and a proofreading polymerase – minimizes the possibility of mutations, offering high yield and fidelity in the PCR.
The optimized RT-PCR Reaction Buffer includes high quality dNTPs and provides the overall improved performance of a hot start system. The buffer′s unique hot start component binds and sequesters primers at lower temperatures to prevent the primers from binding to nonspecific sites. Another component binds magnesium, in a temperature-dependent manner, to prevent uncontrolled DNA synthesis. This new buffer formulation is effective during reverse transcription as well as PCR, resulting in increased specificity and sensitivity.
The combination of optimized reaction buffer and enzyme mix ensures efficient transcription of difficult templates with high secondary structure and GC content without increasing reaction temperature.
The kit′s RT-PCR Enzyme Mix contains four different enzymes: Transcriptor Reverse Transcriptase ensures sensitive and robust reverse transcription with high yield; Protector RNase Inhibitor, fully active at elevated temperatures, provides maximum template protection during reverse transcription; and the Expand System – a blend consisting of Taq DNA Polymerase and a proofreading polymerase – minimizes the possibility of mutations, offering high yield and fidelity in the PCR.
The optimized RT-PCR Reaction Buffer includes high quality dNTPs and provides the overall improved performance of a hot start system. The buffer′s unique hot start component binds and sequesters primers at lower temperatures to prevent the primers from binding to nonspecific sites. Another component binds magnesium, in a temperature-dependent manner, to prevent uncontrolled DNA synthesis. This new buffer formulation is effective during reverse transcription as well as PCR, resulting in increased specificity and sensitivity.
The combination of optimized reaction buffer and enzyme mix ensures efficient transcription of difficult templates with high secondary structure and GC content without increasing reaction temperature.
애플리케이션
The Transcriptor One-Step RT-PCR Kit is designed for fast, sensitive, and specific end-point RT-PCR analysis of RNA (total RNA, mRNA, in vitro-transcribed RNA, or viral RNA) using gene-specific primers.
Featuring an innovative reaction buffer, the kit combines sensitive, high-yield reverse transcription with the improved performance of a hot start system for high-fidelity, high-yield amplification of a variety of templates, including GC-rich RNA.
The kit offers a convenient, highly sensitive one-step RT-PCR method to generate transcripts up to 6.5 kb for applications such as:
The one-step format is convenient, reduces reaction-to-reaction variability, and prevents contamination by minimizing hands-on steps.
Featuring an innovative reaction buffer, the kit combines sensitive, high-yield reverse transcription with the improved performance of a hot start system for high-fidelity, high-yield amplification of a variety of templates, including GC-rich RNA.
The kit offers a convenient, highly sensitive one-step RT-PCR method to generate transcripts up to 6.5 kb for applications such as:
- Qualitative, semi-quantitative, or quantitative analysis of RNA transcription levels
- Determining the presence or absence of RNA templates
- Quantifying gene expression through RNA analysis (e.g., for virus-level quantification)
- Cloning of RNA up to 6.5 kb
The one-step format is convenient, reduces reaction-to-reaction variability, and prevents contamination by minimizing hands-on steps.
Transcriptor One-Step RT-PCR Kit has been used in the amplification of:
- hepatitis C virus (HCV) fragments
- heavy chain CH3 region and light chain polyA region
- murine leukemia virus (MLV)
특징 및 장점
- Obtain high sensitivity and yield in a wide amplification range:
- Achieve high specificity and reduce primer-dimers
- Produce long fragments:
- Transcribe a variety of templates, even the most difficult (e.g., GC-rich RNA):
- Save time and improve results with a convenient kit:
- Protect your RNA sample:
- Increase confidence in your results:
성분
- RT-PCR Enzyme Mix containing Transcriptor Reverse Transcriptase, Protector RNase Inhibitor, and the Expand System
- RT-PCR Reaction Buffer containing hot start mediating components and dNTPs – 5x concentrated
- Water, PCR Grade
- Control RNA – HAV RNA, in vitro transcript *
- Control primer mix HAV RNA (forward and reverse primer) *
* Included only in the 50-reaction pack size.
품질
Each lot of the Transcriptor One-Step RT-PCR Kit is function tested in RT-PCR. RT-PCR is performed with 1000 copies of HAV RNA, in vitro transcript (also provided with Cat.No 04 655 877 001). The control reaction setup and the subsequent RT-PCR with primers for a 246 bp HAV RNA fragment are performed using the standard RT-PCR protocol (reverse transcription at +50°C for 30 minutes) provided in the package insert. With 1,000 copies of target a clearly visible band is obtained after agarose gel electrophoresis and ethidium bromide staining.
In addition, RT-PCR is performed on a dilution series of human liver total RNA (10 ng, 1 ng, and 0.1 ng). RT-PCR with specific primers for a 2.5 kb fragment of ApoB is performed using the standard RT-PCR protocol (reverse transcription at +50°C for 30 minutes) provided in the package insert. With 1 ng of RNA a clearly visible band is obtained after agarose gel electrophoresis and ethidium bromide staining.
In addition, RT-PCR is performed on a dilution series of human liver total RNA (10 ng, 1 ng, and 0.1 ng). RT-PCR with specific primers for a 2.5 kb fragment of ApoB is performed using the standard RT-PCR protocol (reverse transcription at +50°C for 30 minutes) provided in the package insert. With 1 ng of RNA a clearly visible band is obtained after agarose gel electrophoresis and ethidium bromide staining.
기타 정보
For life science research only. Not for use in diagnostic procedures.
유해 및 위험 성명서
예방조치 성명서
Hazard Classifications
Aquatic Chronic 3
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 2
Flash Point (°F)
does not flash
Flash Point (°C)
does not flash
시험 성적서(COA)
제품의 로트/배치 번호를 입력하여 시험 성적서(COA)을 검색하십시오. 로트 및 배치 번호는 제품 라벨에 있는 ‘로트’ 또는 ‘배치’라는 용어 뒤에서 찾을 수 있습니다.
Georg von Massow et al.
Infection and drug resistance, 12, 947-955 (2019-05-24)
Hepatitis C virus (HCV) is a highly variable infectious agent, classified into 8 genotypes and 86 subtypes. Our laboratory has implemented an in-house developed high-resolution HCV subtyping method based on next-generation sequencing (NGS) for error-free classification of the virus using
문서
Understand how mRNA vaccines induce immunity. and read how synthetic mRNA is prepared for vaccine immunogens and other biopharmaceuticals. Find reagents for synthesis of mRNA.
프로토콜
Coronavirus qPCR Primer and probe sets for the detection of SARS-CoV-2 (Corona Virus), with additional real-time RT-PCR, RT-qPCR and supporting reagents available.
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