Fig. 3

Elution condition determination. (a) Effect of elution buffer pH: 100 ng input DNA was used in this experiment. The figure indicates the amount of DNA eluted with low and high pH EBs. A higher amount of DNA was eluted at pH 9.3 compared to pH 8.0 at 74 ⁰C. n = 6 samples were tested for each condition. (b) and (c) Effect of elution temperature: The effect of elution temperature on the amount of DNA eluted was tested using 100 ng (b) and 1000 ng (c) of input DNA. As observed in the panels (b) and (c), 90 ⁰C favored better elution. In experiments in (b) and (c) elution was performed at pH 9.3. n = 5 samples were tested for each condition. (d) Effect of binding method on DNA elution: The graph represents the percentage DNA bound, and the corresponding percentage DNA eluted for three different binding conditions. Binding by ‘orbital shaking’ was tested at 15 min and 2 min. The “tip-based” method was tested for 2 min. For 2 min incubation, “tip-based” method, bound the higher amount of DNA. It also eluted the highest amount of DNA among all three conditions. When “orbital shaking” for 15 min is compared to “orbital shaking” for 2 min, the 2-min sample binds less but elutes higher % DNA. The percent DNA eluted from the total input (100 ng) was calculated to be 27.9, 43.82, and 83.78 for sample incubated with beads for 15 min by orbital shaking, 2 min by orbital shaking, and 2 min by “tip-based” method, respectively. The error bar in the graph depicts the SD. n = 6 for each condition.