molecular biology

end-point PCRSuperScript ® III Reverse TranscriptaseEngineered for higher thermostability, increased half-life, and reduced RNase H activity• Half-life of 220 minutes at 50°C for high cDNA yields• Reduced RNase H activity for more full-length cDNA• Full activity at 50°C for increased specificity with gene specificprimers (GSPs)• Ability to increase RT units without inhibiting subsequent PCRkb9.57.5SuperScript ® IIIM 37˚ 42˚ 45˚ 50˚ 55˚SuperScript ® II M-MLV37˚ 42˚ 45˚ 50˚ 55˚ 37˚ 42˚ 45˚ 50˚ 55˚ MSuperScript ® III Reverse Transcriptase (RT) is a proprietary mutantof SuperScript ® II RT that is active at 50°C and has a half-life of 220minutes, providing increased specificity with gene specific primersand high cDNA yield. Like SuperScript ® II RT, it synthesizes a complementaryDNA strand from single-stranded RNA, DNA, or an RNA:DNAhybrid. SuperScript ® III RT can be used for RT-PCR of a specific gene,or for generating cDNA from total or poly (A)+ RNA samples. It is idealfor synthesis of first-strand cDNA, array labeling, cDNA libraries,RT-PCR, primer extension, and 3´ and 5´ RACE.4.42.41.4Comparison of SuperScript ® III Reverse Transcriptase toSuperScript ® II RT and M-MLV RT. An autoradiograph isshown of 32 P-labeled cDNA synthesized from a mixture of0.25 µg of RNA each of 1.35 kb, 2.4 kb, 4.4 kb, 7.5 kb, and9.5 kb with 200 units of each RT at various temperatures.Lane M is 32 P-labeled 1 kb DNA ladder.Product Quantity Cat. No.SuperScript ® III Reverse Transcriptase 10 reactions 18080-09350 reactions 18080-044200 reactions 18080-085SuperScript ® III First-Strand Synthesis SystemHigh cDNA yields in a convenient kit format• Contains SuperScript ® III ReverseTranscriptase for higher cDNA yields• Detects template molecules from aslittle as 1.0 pg total RNA• Includes all components needed forfirst-strand cDNA synthesis• Consists of optimized reagents formore robust performanceTotal HeLaRNA (ng)FIB 9.4 kbPol ε 6.8 kbVIN 4.6 kbMPromegaImProm-II RT System42˚C – 60 min1 100MStratageneProSTAR ®First StrandRT-PCR Kit42˚C – 60 min1 100Applied Biosystems ®GeneAmp ® GoldRNA PCR Kit42˚C – 12 min1 100MRocheFirst Strand cDNASynthesis Kit forRT-PCR (AMV)42˚C – 60 min1 100MMInvitrogen SuperScript IIIFirst-StrandSynthesisSystem for RT-PCR50˚C – 50 min1 100M3RT-PCRThe SuperScript ® III First-StrandSynthesis System for RT-PCR providesall the necessary components forsynthesis of high-quality, first-strandcDNA from total or poly(A)+ RNA.Used with PCR, this functionally testedsystem enables you to detect raremessages and generate enough materialfor cloning.PP2A 1093 bpFigure 2. The SuperScript III First-Strand Synthesis System outperforms the competition.RT reactions containing 1 and 100 ng of total HeLa RNA were performed with each kit usingreagents and conditions specified in each manufacturer’s protocol. Ten percent (2-5 μL) ofthe resulting cDNA was added to PCR reactions containing 1 unit of Platinum ® Taq DNAPolymerase High Fidelity for 35 PCR cycles, 1 min/kb. PCR products were separated on a1% agarose gel containing 0.4 μg/mL ethidium bromide.1_2.tiffProduct Quantity Cat. No.SuperScript ® III First-Strand Synthesis System 50 reactions 18080-051www.lifetechnologies.com146