Invitrogen™ RETROscript™ Kit with Manual

Beschreibung

• Amplify specific RNA targets from complex mixtures (i.e., total cellular RNA)
• Both random decamers and oligo(dT)₁₈ primers provided
• Perform either one- or two-step RT-PCR
• Sufficient for 40 reactions

Using the RETROscript Kit
The RETROscript Kit provides reagents for reverse transcription of cDNA from RNA templates. When used with SuperTaq thermostable DNA polymerase (available separately), the cDNA templates are efficiently amplified into double-stranded DNA products. The molony-murine leukemia virus (M-MLV) reverse transcriptase is used in the first step to extend a primer hybridized to an RNA sample. M-MLV RT has higher stability and lower intrinsic RNase H activity than AMV reverse transcriptase. Both random decamers and oligo dT are provided in the RETROscript Kit. The kit also includes two alternative buffers for first strand synthesis. Protocols are provided for performing the reverse transcription and PCR amplification reactions sequentially (two-step) or as a single reaction. Optimal reaction temperature: 44° to 55°C.

Accessory products
SuperTaq Polymerase (Mfr. Nos. AM2054 and AM2056) and SuperTaq Plus Polymerase (Mfr. Nos. AM2054 and AM2056) are recommended for use with the RETROscript Kit. RETROscript Kit components that are available separately include dNTPS for cDNA Probe Synthesis (Mfr. No. AM8200), Oligo (dT) Primer (Mfr. No. AM5730G), and Random Decamers (Mfr. No. AM5722G). Also available are the T7-Oligo (dT)₂₄ Promoter Primer (Mfr. No. AM5710) and the T7-Oligo (dT)₂₄ Promoter Primer, Anchored (Mfr. No. AM5712).

PCR & Real-Time PCR, Reverse Transcription