TABLE 2.

Real-time PCR analysis of effects of histone alleles on expression of telomeric ADE2^a

Substitution	ΔΔCt (+/−SD)	Relative expression of subtelomeric ADE2
WT H4	0	1
LTS allele	−1.36 (0.11)	2.6
H4 K16A	−1.32 (0.16)	2.5
H4 K16R	0.34 (0.15)	0.8
H4 K16Q	−1.85 (0.13)	3.6
H4 K77A	−1.31 (0.09)	2.5
H4 K77R	1.14 (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

^aRNA 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. ΔΔC_t was calculated using the formula (C_t ADE2 − C_t ACT1)_mutant − (C_t ADE2 − C_t 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.