使用Transcriptor First Strand cDNA Synthesis Kit在对各种丰度的RNA进行高效反转录，生成的cDNA产物真实体现起始样本的基因表达水平。配合使用LightCycler®480或其他RT-PCR仪器以及基因特异引物、通用文库探针文库、SYBR Green I，通过qPCR扩增目标基因。
该试剂盒包含了用于逆转录反应的所有试剂，包括独特的锚定寡核苷酸(dT)18引物，RNase抑制剂，对照RNA以及对照PBGD引物混合物，满足您cDNA合成全部的需要。
■ 提升RT-PCR反应灵敏度
同时反转录低丰度及高丰度RNA，提升反应的灵敏度。代表起始样本真实基因表达水平的cDNA合成产物，确保后续的qPCR反应的准确性。
■ 能够对108范围内的RNA进行精确的线性定量无论是对低表达水平基因还是高表达水平基因进行分析，皆得心应手。
■ 简化结果分析
生成的平滑qPCR曲线，呈现更强的荧光信号，且能保证不同RNA稀释梯度之间的均一性。
■ 克服复杂二级结构
高达55℃的逆转录温度，轻松转录高GC含量模板。
■ 获取大量全长cDNA
最长可轻松合成长达14kb的全长cDNA
产品说明:
- 锚定引物——Oligo (dT)18，让cDNA合成从poly(A)的5'端开始，减少无效延伸，提供反应效率。
- 试剂盒同时提供了六聚体随机引物。
- 50次反应规格的试剂盒同时提供了10次反应的阳性对照。
灵敏度更高 - 对样本中的低丰度或高丰度RNA可同时进行转录，确保最初的基因表达水平不被改变。
RNA模板适用性更广 - 55ºC条件下，可高效完成对带有复杂二级结构或富含GC的RNA模板的反转录。
反应速度更快 - 仅需10分钟就能合成全长cDNA。
全长cDNA产量更高 - 试剂盒配备独有的锚定oligo(dT)18 引物，确保获取真正的全长cDNA，最长可转录达14 kb的转录子。
准确性最高 - Transcriptor High Fidelity cDNA Synthesis Kit提供了同类产品中独有的准确度保证；较之常规的转录酶，保真度提升7倍！
试剂盒组成:
- 逆转录酶：Transcriptor Reverse Transcriptase
- 逆转录酶缓冲液：Transcriptor RT Reaction Buffer, 5x concentrated
- RNase抑制剂：Protector RNase Inhibitor
- dNTP Mix, PCR Grade
- 锚定引物：Anchored Oligo (dT)18 Primer
- 六聚体随机引物：Random Hexamer Primer
- Control RNA – total RNA fraction purified from the immortalized cell line K-562.
Note: Included only in the 50-reaction pack size - Control PBGD Primer Mix (forward and reverse primer)
Note: Included only in the 50-reaction pack size
- PCR级水：Water, PCR Grade
产品应用：
The Transcriptor First Strand cDNA Synthesis Kit is designed to reverse transcribe RNA (mRNA, total RNA, viral RNA, and in vitro-transcribed RNA) from a variety of sources for the following applications:
- Studying gene expression levels, via two-step RT-PCR, using qualitative RT-PCR or quantitative RT-PCR on the LightCycler® Carousel-Based System, the LightCycler® 480 System, or other real-time instruments.
- Generating cDNA libraries with large and full-length inserts.
- Cloning genes of interest.
The kit contains all components required for cDNA reactions for use with conventional thermal cyclers and real-time PCR instruments. In addition, the 50-reaction pack size includes 10 control reactions.
产品特点：
Optimized for Many Applications
- Use with conventional thermal cyclers and real-time PCR instruments.
- Optimize real-time PCR – the kit is tested with the LightCycler® Carousel-Based System, the LightCycler® 480 System, and other real-time instruments.
- Obtain accurate linear quantification over at least a 108 -fold range of input RNA (in vitro transcripts), permitting the analysis of genes with very low or extremely high expression levels.
- Generate efficient qPCR curves, with high fluorescence intensity and identical distances between RNA dilutions, allowing a straightforward analysis of results (Figure 1).
- Obtain reliable and sensitive results – there are no additives in the RT buffer system that interfere with – or inhibit – the subsequent PCR.
Robustness
- Transcribe a variety of templates, even the most difficult (e.g., GC-rich RNA with high secondary structures).
- Achieve high reproducibility (Figure 2).
- Generate full-length transcripts with the anchored-oligo(dT)18 primer that is included in the kit.
- Obtain cDNA transcripts up to 14 kb (Figure 3).
Flexibility
- Use three different priming methods – random hexamers, anchored-oligo(dT)18, and sequence-specific primers – depending on the type of analysis needed.
产品描述：
Transcriptor Reverse Transcriptase – the core component of the kit – is a recombinant reverse transcriptase expressed in E. coli. The enzyme has RNA-directed DNA polymerase activity, DNA-dependent DNA polymerase activity, unwinding activity, and RNase H activity that degrades RNA in RNA:DNA hybrids. The latter circumvents the need to perform an additional time-consuming RNase H incubation step after reverse transcription. This shortens the reaction time and reduces costs.
Transcriptor Reverse Transcriptase is recommended for RT-PCR because of its high sensitivity in combination with high thermostability: the enzyme synthesizes long cDNA products (up to 14 kb) and can be used at temperatures up to 65°C. Due to its thermostability, Transcriptor Reverse Transcriptase is recommended for GC-rich templates with high secondary structure, without the need to include additives in the reaction.
The kit provides all reagents required for first-strand cDNA synthesis reactions. For priming, three different primer systems can be used. Two cDNA synthesis primers are provided with the kit: random hexamer primers and an anchored-oligo(dT)18 primer. The latter is designed to bind at the beginning of the poly(A) tail to generate full-length cDNAs and to prevent priming from internal sites of the poly(A) tail. The 5' ends of long mRNAs are often underrepresented; therefore, this priming method is preferred for most applications. The use of random hexamer primers enables priming throughout the length of RNA for uniform representation of all RNA sequences and allows reverse transcription of RNAs that do not carry a poly(A) tail.
Thermostable Protector RNase Inhibitor is included in the kit to protect RNA from degradation at high reaction temperatures.
背景信息：
Reverse Transcriptase
Transcriptor Reverse Transcriptase – the core component of the kit – is a highly sensitive, highly thermostable, recombinant reverse transcriptase. The enzyme has RNA-directed DNA polymerase activity, DNA-dependent DNA polymerase activity, unwinding activity, and RNase H activity (for degrading RNA in RNA:DNA hybrids). The presence of the latter activity circumvents the need for an additional time-consuming RNase H incubation step after reverse transcription. Thus, Transcriptor Reverse Transcriptase is an ideal enzyme for first strand cDNA synthesis.
The enzyme can be used at temperatures up to 65 C. Due to its thermostability, Transcriptor Reverse Transcriptase can accurately transcribe GC-rich templates that have large amounts of secondary structure. This property of Transcriptor Reverse Transcriptase eliminates the need for special reaction-enhancing additives.
Random hexamer primers
Random hexamer primers (included in the kit) bind throughout the entire length of RNA, ensuring reverse transcription of all RNA sequences. These primers also allow reverse transcription of RNAs that do not carry a poly(A) tail.
Anchored oligo(dT)18 primer
The anchored-oligo(dT)18 primer (included in the kit) is designed to bind at the beginning of the poly(A) tail (rather than randomly within the tail) to generate full-length cDNAs. Since the 5' ends of long mRNAs are often underrepresented in cDNA mixtures, this primer is preferred for most applications.
Protector RNase Inhibitor
Thermostable Protector RNase Inhibitor (included in the kit) protects RNA from degradation at high reaction temperatures.
订货信息：
| Product | Transcriptor First Strand cDNA Synthesis Kit |
|---|---|
| Company | Roche Applied Science |
| Price |
询价
|
| More Information | |
| Catalog Number | 4379012001 |
| Quantity | 1 kit for up to 50 reactions, including 10 control reactions |
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