Ambion, Inc - Products

RETROscript® First Strand Synthesis Kit for RT-PCR

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	Cat#	Product Name	Size
	AM1710	RETROscript® Kit	40 rxns
	AM8200	dNTPs for cDNA Probe Synthesis (10 mM)	4 x 75 µl
	AM5730G	Oligo (dT) Primer (50 µM)	0.08 ml
	AM5722G	Random Decamers (50 µM)	0.08 ml
NEW!	AM1710M	RETROscript® Kit w/Manual	40 rxns
	AM2054	SuperTaq™ Plus Polymerase (Cloned) 5 U/µl	50 U
	AM2056	SuperTaq™ Plus Polymerase (Cloned) 5 U/µl	250 U
	AM2050	SuperTaq™ Polymerase (Cloned) 5 U/µl	50 U
	AM2052	SuperTaq™ Polymerase (Cloned) 5 U/µl	250 U
	AM5712	T7-Oligo(dT)₂₄V Promoter Primer, Anchored (100 ng/µl)	40 rxns
For Research Use Only. Not for use in diagnostic procedures.

Amplify specific RNA targets from complex mixtures (i.e. total cellular RNA)
Both random decamers and oligo(dT)₁₈ primers provided
Complete Instruction Manual with protocols for both one- and two-step RT-PCR
SuperTaq™ thermostable DNA polymerase available separately

Ambion's RETROscript First Strand Synthesis 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. Total RNA, poly(A) RNA, tRNA or rRNA from a variety of sources (e.g. bacterial, plant, yeast) can be used as a template for reverse transcription. With RETROscript, it is usually not necessary to isolate the polyadenylated fraction, even for amplification of rare messages.

The Molony-Murine Leukemia Virus (M-MLV) Reverse Transcriptase is used in the first step to extend a random decamer, oligo dT or specific primer hybridized to an RNA sample containing the message of interest. We have chosen M-MLV for its higher stability and lower intrinsic RNase H activity, compared to AMV Reverse Transcriptase. Both random decamers and oligo dT are provided in the RETROscript Kit to allow for the option of amplifying full length cDNA when desired. Note that one or the other type of primer may be better for maximizing yield of an amplified mRNA depending on the specific target (e.g. compare amplification of beta-actin versus S15). A pair of specific primers provided by the researcher is then used to amplify the target region via PCR.

Ambion's RETROscript Kit provides protocols and reagents for performing the reverse transcription and PCR amplification reactions sequentially (two-step) or as a single reaction. The kit also includes two alternative buffers for first strand synthesis: the 10X RT-PCR Buffer may be used for both the reverse transcription and the amplification reactions, in either a two-step or single-step reaction, while the Alternate 10X First-Strand Buffer may be used to maximize the total yield of cDNA made with M-MLV reverse transcriptase. A positive control target and primers allow troubleshooting of the kit reagents and procedure. These primers also often permit troubleshooting of the user's template RNA sample since the control primers are designed to amplify a region of a conserved, constituitively expressed gene (the ribosomal protein S15 gene). The kit contains a High Resolution Gel Loading Solution that is especially useful for assessing short products (<500 bp). A comprehensive Instruction Manual provides background information as well as information on related procedures (e.g. producing templates for in vitro transcription using RT-PCR).

SuperTaq and SuperTaq Plus are trademarks of and are manufactured by Enzyme Technologies Ltd. and sold under licensing arrangements with F. Hoffman-La Roche, Ltd., Roche Molecular Systems, Inc. and the Perkin-Elmer Corporation. Ambion, Inc. is a distributor of Enzyme Technologies, Ltd., except in the following countries: Austria, Benelux, Denmark, Greece, Sweden, Switzerland, Taiwan, and the United Kingdom.

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