Half-life calculation
RNA half-lives can be calculated with HALO using an exponential decay model either for individual
replicates or ratios averaged across all replicates. Three possible methods exist for the calculation
of half-lives, based on the newly transcribed/total RNA, pre-existing/total RNA and the newly transcribed/pre-existing RNA ratios,
respectively. Among these, the method based on newly transcribed (=labeled) to total RNA ratio has an overall high
accuracy, while the method using pre-existing/total RNA shows high precision for short transcripts, while
being unreliable for medium-to-long-lived transcripts (Friedel
and Döken, 2008). The ratio of newly transcribed to pre-existing RNA as a method
combines the good properties of the two other calculation methods.