Skip to main content
. Author manuscript; available in PMC: 2012 Sep 23.
Published in final edited form as: J Mol Biol. 2011 Jul 29;412(3):354–364. doi: 10.1016/j.jmb.2011.07.055

Figure 3. Strand transfer with E478Q mutant RT using PCIS substrates.

Figure 3

(a) A 10% denaturing gel image of time course reactions performed with E478Q mutant RT using the substrate without a pre-created invasion site (PCIS) (see Figure 1a) and (b) gel image of reactions with a PCIS substrate (Figure 1b). A master reaction was prepared containing primer-donor, acceptor, magnesium and E478Q mutant RT. After pre-incubation for three minutes at 37°C, the reaction was started by adding dNTPs. Aliquots were then taken and mixed with stop buffer after reaction times of 30 sec, 1, 3, 5, 10 and 15 min. TP denotes transfer product, DE, donor extension product and P, unextended primer. Panel (c) represents analysis of strand transfer products: shown on the Y-axis, % transfer efficiency, while on the X-axis are products analyzed on each time point using the substrate without (−) PCIS or with (+) PCIS.