Reproducibility and sensitivity of the pro MSS HTA.
(A) The MSS HTA reflects the populations in the PCR
product accurately and reproducibly at high template numbers. PCR
products of two pro genes were mixed at known ratios;
the mixtures were then diluted over a 100-fold range and annealed to
the MSS HTA probe 6.1. The abundance of one of the products was
determined by using the MSS HTA and compared with the known abundance.
Error bars represent standard deviations from 5–9 experiments.
(B) Amplification of pro gene mixtures.
Mixtures of pro genes were amplified and quantified by
using the MSS HTA. The smallest number of templates required to detect
and quantify a 10% population reproducibly is 1,200. Error bars are
the standard deviation from three experiments. The sequences used
encoded the following changes from Hxb: (1) T12S, K43R, M46I, Q61L,
L63P, V82A; (2) T12S, K43R, M46I, I54V, Q61H, L63P, V82F.