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反轉(zhuǎn)錄酶III 逆轉(zhuǎn)錄酶三 invitrogen*原裝 SuperScript® III Reverse Transcriptase
貨號18080-044
與SuperScriptII逆轉(zhuǎn)錄酶一樣,SuperScriptIII的RNase H 活性較低,能獲得更高產(chǎn)量的全長Cdna。另外,它還有著其它的逆轉(zhuǎn)錄酶所無法提供的熱穩(wěn)定性;經(jīng)過基因工程改造的SuperScriptIII具有更高的熱穩(wěn)定性,其半衰期在50度時達到令人難以置信的220分鐘,這個特性也提高SuperScriptIII轉(zhuǎn)錄具有二級結(jié)構(gòu)模板RNA的能力。
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invitrogen:www.invitrogen。。com
SuperScript® III Reverse Transcriptase (RT) is a proprietary mutant of SuperScript® II RT that is active at 50°C and has a half-life of 220 minutes, providing increased specificity with Gene-Specific Primers (GSPs) and the highest cDNA yield of all RTs. It is ideal for RT-PCR of a specific gene or generating cDNA from total or poly (A)+RNA sample. Like SuperScript® II, it synthesizes a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. SuperScript® III RT is genetically engineered by the introduction of point mutations that increase half-life, reduce RNase activity, and increase thermal stability.
•Thermostability– Half life of 220 minutes at 50°C
•Yield– Reduced RNase H activity for more full-length cDNA
•Specificity– Full activity at 50°C for increased specificity with Gene Specific Primers
•Applications– Array labeling, cDNA libraries, RT-PCR, primer extension, and 3´ and 5´ RACE
Source: Purified fromE. coliexpressing thepolgene of M-MLV (1, 2), mutagenized to increase thermal stability and half life and reduce RNase H activity.
Performance and Quality Testing: SDS-PAGE purity; endodeoxyribonuclease, exodeoxyribonuclease, and ribonuclease assays (Figure 2) and yield and length of cDNA product.
Unit Definition: One unit of SuperScript® III RT 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)12-18as a 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.
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