In vitro transcription with T7 RNA polymerase: Difference between revisions

In vitro transcription with T7 RNA polymerase (view source)

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 '''1X buffer:'''
-mM Tris-Cl, pH 7.5
+mM Tris-HCl, pH 7.5
-mM MgCl2 (How do you make superscripts and subscripts?)
+mM MgCl<sub>2
 mM dithiothreitol (DTT)
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 Inorganic pyrophosphatase (PPase) catalyzes the hydrolysis of inorganic pyrophosphate to form orthophosphate:
-P2O7-4 + H2O -> 2HPO4-2. Inorganic pyrophosphate is released when a nucleoside triphosphate is incorporated/polymerized into the growing chain. This helps to prevent against any inhibitory effect of having pyrophosphate around (i.e. prevents the "reverse" reaction.) This is an optional component of the transcription reaction. If you leave it out, often you will see something precipitate (white) in your transcription reaction. This is the pyrophosphate.
+P<sub>2</sub>O <sub>7 <sup>-4 + H2O -> 2HPO<sub>4 <sup>-2. Inorganic pyrophosphate is released when a nucleoside triphosphate is incorporated/polymerized into the growing chain. This helps to prevent against any inhibitory effect of having pyrophosphate around (i.e. prevents the "reverse" reaction.) This is an optional component of the transcription reaction. If you leave it out, often you will see something precipitate (white) in your transcription reaction. This is the pyrophosphate.
 Make a 0.1 U/µL stock solution in H2O and store at -20 ˚C.