The Large Scale Transcription Kits are designed for convenient and consistent in vitro production of large amounts of RNA from sequences cloned in vectors containing T7 or SP6 promoters. The transcription conditions have been optimized to provide maximal yields of full-length RNA that can be used in a variety of applications, including in vitro translation, structural analysis, splicing, antisense, and ribozyme synthesis. Under these conditions, more than 700 moles of transcript can be synthesized per mole of template (Noren 1990).

The nucleoside triphosphates are provided in separate vials to allow substitution of modified or labeled NTPs under limiting conditions for synthesis of probes. The kits also include a positive control template to monitor the performance of test DNAs. The kits include enough reagents to perform 100 synthesis reactions (100 µl volume), each of which will produce more than 100 µg of RNA from 5 μg template in 60 minutes under non-limiting conditions. Also, the bacteriophage T7 and SP6 RNA polymerases and RNase-free DNase I included in the kits are available separately.

The Large Scale SP6 Transcription Kit is designed for convenient and consistent in vitro production of large amounts of RNA from sequences cloned in vectors containing the SP6 promoter. The transcription conditions have been optimized to provide maximal yields of full-length RNA that can be used in a variety of applications, including in vitro translation, structural analysis, splicing, anti-sense, and ribozyme synthesis. Under these conditions, more than 700 moles of transcript can be synthesized per mole of template (1). The nucleoside triphosphates are provided in separate vials to allow substitution of modified or labeled NTPs under limiting conditions for synthesis of probes. The kits also include a positive control template to monitor the performance of test DNAs. The kits include enough reagents to perform 100 synthesis reactions (100 µl volume), each of which will produce more than 50 µg of RNA from 5 µg template in 60 minutes under non-limiting conditions. •;-;10,000 units;-;T7 RNA Polymerase or SP6 RNA Polymerase
•;-;1 ml;-;20 mM ATP
•;-;1 ml;-;20 mM CTP
•;-;1 ml;-;20 mM GTP
•;-;1 ml;-;20 mM UTP
•;-;2 × 1 ml;-;5X Transcription Buffer
•;-;1 ml ;-;3 M Na Acetate, pH 5.2
•;-;100 µl;-;1 M DTT
•;-;4 × 1.5 ml;-;Nuclease-free Water
•;-;1000 U;-;RNase-free DNase I
•;-;5 µg;-;Control Template