RETROscript® Kit with Manual

Catalog number: AM1710M

Invitrogen™  Related applications: Reverse Transcription

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Description

The RETROscript® Kit includes reagents for reverse transcription of RNA into double-stranded cDNA products. Advantages of the RETROscript® Kit:

• 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

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.

Accessory products
SuperTaq™ Polymerase (Cat. Nos. AM2054 and AM2056) and SuperTaq™ Plus Polymerase (Cat. 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 (Cat. No. AM8200), Oligo (dT) Primer (Cat. No. AM5730G), and Random Decamers (Cat. No. AM5722G). Also available are the T7-Oligo (dT)24 Promoter Primer (Cat. No. AM5710) and the T7-Oligo (dT)24 Promoter Primer, Anchored (Cat. No. AM5712).
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Reverse Transcriptase: M-MLV Reverse Transcriptase
Downstream Application: Reverse Transcriptase PCR (RT-PCR)
Reaction Format: Separate Components
Sample Type (General): RNA
Product Size: 40 reactions
Final Product: cDNA
Optimal Reaction Temperature: 44-55° C

Contents & storage

• 50 µl Control Template RNA (0.5 mg/ml)
• 200 µl 10X RT Buffer
• 400 µl 10X PCR Buffer
• 300 µl dNTP mix (2.5 mM each dNTP)
• 80 µl Random Decamers (50 µM)
• 80 µl Oligo(dT) Primers (50 µl)
• 40 µl MMLV-RT (100 units/µl)
• 40 µl RNase Inhibitor (10 units/µl)
• 50 µl Control PCR Primers (5 µM each)
• 1.0 ml High Resolution Gel Loading Solution (5X)
• 3.5 ml Nuclease-free Water

Nuclease-free Water may be stored at any temperature. Store Control Template RNA below -70°C. Store all other reagents at -20°C.

Documents

Manuals & protocols

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