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. 2007 Aug;13(8):1384–1389. doi: 10.1261/rna.528007

FIGURE 2.

FIGURE 2.

(A) Ethidium bromide stained 12% denaturing polyacrylamide gel of transcription of a 94-nt RNA with the 3′ affinity tag attached in different transcription buffers. (Lane 1) Standard Tris transcription buffer with 20 mM DTT; (lane 2) standard Tris transcription buffer with 10 mM DTT; (lane 3) transription with Na-HEPES (pH 8.0) and 20 mM DTT; (lane 4) transcription with Na-HEPES (pH 8.0) and 10 mM DTT. Product “a” is the full-length product, “b” is the cleaved 3′-tag, and “c” is the 94-nt desired product RNA (which is also the size marker in the “MW” lane). (B) 4%–12% SDS-PAGE analysis of expression and purification of the HMM protein. (Lane 1) Soluble fraction of total cellular lysate; (lane 2) elution from the nickel affinity column; (lane 3) peak from the SP-column. (*) The product protein; molecular size standards are to the right of the gel.