Berglund:Transcription Capped

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In vitro Transcription with ultrahot/capped RNA for splicing

5µl 5xT7 buffer
1µl 400mM DTT
2.5µl rNTP Mix (5mM A, U, 1mM G)
2.5µl 0.1mM CTP
5µl P32 CTP
1µl 10mM M7cap
0.5µl Rnasin
0µl water
5µl plasmid or PCR (1µg)
2µl T7 polymerase
2.5µl 10xyeast pyrophosphatase
25µl total


1x buffer:
40mM Tris pH 8
6mM MgCl2
2mM spermidine
10mM NaCl
25mM Tris pH 9