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. 2008 Aug 22;36(18):e121. doi: 10.1093/nar/gkn531

Figure 2.

Figure 2.

Purified polymerases with AviTag legs. (T) Therminator protein purchased from New England Biolabs. (P) The parent enzyme (a mutant of Therminator selected for improved utilization of phosphate-labeled dNTPs) has a higher molecular weight compared to Therminator because it has a C-terminal Myc-His fusion used for purification. (B53) Polymerase ‘P’ with AviTag leg at position K53. (B229) Polymerase ‘P’ with AviTag leg at position 229. (DBio; ‘dual biotin’) Polymerase ‘P’ with two AviTag insertions at K53 and K229. Duplicate samples (5 μl each) of the Ni-NTA purified proteins were resolved by SDS–PAGE [Invitrogen NuPAGE Bis–Tris (MOPS) 4–12%]. The gel was stained with Coomassie blue and imaged with a LI-COR Odyssey infrared imager (application note http://www.licor.com/bio/Odyssey2/Odyssey13.jsp). Marker sizes (kDa) are indicated.