Figure 7

Real time qRT-PCR analysis of FLC expression on plant samples harvested after four-weeks cold treatment and extracted using the HTP96 method. Reverse transcription was carried out with and without RNA adjustment. Reverse transcription (RT) was simplified by using a defined volume of RNA preparation in each RT reaction. Plants samples of four different accessions were collected ten days (4V10), 24 days (4V24) or 30 days (4V30) after a four-week cold treatment. Total RNA was extracted using the HTP96 method. The reverse transcription reaction was performed using either 2 μL of RNA preparation, or by adjusting RNA concentration to 1 μg in a 10 μL final reaction volume. Relative FLC expression was assayed by SYBR green real time qRT-PCR using three technical replicates. Error bars represent the standard error of the mean. A. FLC expression assayed without prior RNA quantification. B. FLC expression assayed from 1 μg of total RNA.