32 transcripts is an optimum threshold for achieving similar cumulative frequency distribution amongst replicates. KS statistics was applied iteratively to our biological replicates (red and black lines), each graph depicts a gradual change in the threshold value. The KS test statistics can be thought of as a cost function that we seek to minimize to ensure that the distributions between the two replicates are similar. Applying such a strategy iteratively as we change the threshold value gradually, we arrive at a point where both line converge together indicating similar distributions. The point where this first occurs is designated as the minimum threshold count value of biological significance.