With the MEGAscript T7 Transcription Kit, can the T7 polymerase synthesize RNA from a single-stranded DNA template, containing the T7 promoter sequence, or does it require a double-stranded DNA promoter sequence to work?
The T7 polymerase requires, at the minimum, for the promoter sequence to be double-stranded. The rest of the template does not necessarily have to be double-stranded.
Is there a minimum length of template that can be used with the MEGAscript kit?
The MEGAscript Kit is designed to function best with templates that code for RNA transcripts of about 0.5 kb and longer. The kit can be used to produce shorter RNA, but modify the reaction as described in section Optimizing yield of short transcripts on page 24 of the manual.