Real-time PCR analysis of effects of histone alleles on expression of telomeric ADE2a

SubstitutionΔΔCt (+/−SD)Relative expression of subtelomeric ADE2
WT H401
LTS allele−1.36 (0.11)2.6
H4 K16A−1.32 (0.16)2.5
H4 K16R0.34 (0.15)0.8
H4 K16Q−1.85 (0.13)3.6
H4 K77A−1.31 (0.09)2.5
H4 K77R1.14 (0.18)0.4
H4 K77Q−0.50 (0.02)1.4
H4 K79A−1.0 (0.37)2
H4 K79R−0.68 (0.01)1.6
H4 K79Q−1.07 (0.41)2.1
  • a RNA was prepared from the indicated substitution strains and reverse transcribed into cDNA. Quantitative PCR was undertaken to amplify both ADE2 and ACT1 cDNA in the presence of SYBR green. ΔΔCt was calculated using the formula (Ct ADE2 − Ct ACT1)mutant − (Ct ADE2 − Ct ACT1)wild type. Values were also compared with those of an RT-negative control for each sample. Results are representative of RNA that was prepared from two individual colonies of each sample, both of which were assayed in quadruplicate.