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- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 保存条件:
低温
- 库存:
现货
- 供应商:
上海善然生物科技有限公司
- 规格:
2,000 units
描述
SuperScript® II Reverse Transcriptase (RT) is an improved version of SuperScript® RT. It is a DNA polymerase that synthesizes a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. This enzyme is genetically engineered by the introduction of point mutations in the RNase H active center to reduce RNase H activity. This structural modification eliminates degradation of RNA molecules during first-strand cDNA synthesis and gives SuperScript® II RT superior performance characteristics.• Yield – Reduced RNase H activity results in greater first-strand cDNA yields
• Size of cDNA – Generation of RT-PCR products up to 12 kb
• Thermostability – Full activity at 42°C
• Flexibility – Uses either total or poly(A)+ RNA
Performance and Quality Testing: SDS-PAGE purity; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays; and yield and length of cDNA product.
Unit Definition: One unit of SuperScript® II is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min. at 37°C using poly(A)•oligo(dT)12-18 as template•primer (3).
Unit Reaction Conditions: 50 mM Tris-HCl (pH 8.3), 40 mM KCl, 6 mM MgCl2, 1 mM DTT, 0.5 mM [3H]dTTP, 0.1 mM poly(A), 0.1 mM oligo(dT)12-18, 0.1 mg/ml BSA, and enzyme in 50 µl for 10 min. at 37°C.
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.
规格
| Optimal Reaction Temperature: | 42° C |
|---|---|
| Final Product Size(s): | 12.3 kb or less |
| Reverse Transcriptase: | SuperScript® II |
| Ribonuclease H Activity: | Reduced |
| Form: | Frozen |
| Format: | Tube(s) |
| Quantity: | 2000 units |
| Template: | ssDNA, ssRNA, RNA-DNA Hybrid |
| Sensitivity: | Medium |
| Product Size: | 2,000 units |
| Concentration: | 200 U⁄µl |
| Enzyme Function: | RNA dependent DNA Polymerase |
| Shipping Condition: | Wet Ice |
| Number of Reactions: | 10 Reactions |
内容及储存
Store at -20°C. Guaranteed stable for 6 months when properly stored.
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文献和实验cDNA synthesis using superscript II
SuperScript II 1 µl water 4 µl Prepare this mix as a master mix and add then 20 µl to each reaction. Synthesize the cDNA in a thermocycler: 1.
SuperScript III One-Step RT-PCR System with Platinum® Taq High Fidelity
实验原理 The system consists of two major components: SuperScript™ III RT/ Platinum® Taq High Fidelity Enzyme Mix and 2X Reaction Mix. SuperScript™ III Reverse Transcriptase is a version of M-MLV RT that has been engineered
分离高质量RNA 成功的cDNA合成来自高质量的RNA。高质量的RNA至少应保证全长并且不含逆转录酶的抑制剂,如EDTA或SDS。RNA的质量决定了你能够转录到cDNA上的序列信息量的最大值。一般的RNA纯化方法是使用异硫氰酸胍/酸性酚的一步法。Trizol试剂法(见下图)将一步法加以提高,可以从多种组织和细胞中提取高质量的非降解RNA。Trizol试剂法可以从最少100个细胞或1mg组织中提取RNA。TRIzol®试剂步骤略图 一般不必使用oligo(dT)选择性分离poly
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