We strongly recommend wearing gloves and using nuclease-free tubes and reagents to avoid RNase contamination. Reactions are typically 20 μl but can be scaled up as needed. Reactions should be assembled in nuclease-free microfuge tubes or PCR strip tubes.
The kit formulation allows for efficient capped RNA synthesis using cap analog
(ARCA). The recommended ratio of cap analog to GTP is 4:1. Increasing
the ratio of cap analog to GTP will increase the proportion of capped RNA
transcripts; however, it also significantly decreases the yield of the reaction.
Cap analogs are sold separately. Please refer to the companion products section.
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The NEB PCR Cloning Kit allows direct cloning from any amplification reaction, with no purification needed!