Table 2.

PLS Regression Analysis

X variablesResults of SelectionResults of first (1) and second (2) optimization 1 and 2 (180 nt)1 and 2 (360 nt)
180 nt360 nt1 (180 nt)1 (360 nt)2 (180 nt)2 (360 nt)
Ligation temp.−0.45 ± 0.16−0.23 ± 0.11−0.64 ± 0.02−0.41 ± 0.06NS NS −0.22 ± 0.04−0.18 ± 0.02
Cycles0.35 ± 0.160.28 ± 0.110.17 ± 0.050.34 ± 0.070.077 ± 0.040.046 ± 0.02
Enzyme conc.NS 0.24 ± 0.110.20 ± 0.070.32 ± 0.070.10 ± 0.09NS 0.088 ± 0.04
DNA conc.0.22 ± 0.16NS NS NS
Oligo conc.NS NS
Ligation timeNS NS 0.19 ± 0.050.22 ± 0.06−0.078 ± 0.04−0.12 ± 0.02
Temp. (2)0.32 ± 0.050.22 ± 0.16−0.24 ± 0.07−0.25 ± 0.08−0.13 ± 0.02−0.081 ± 0.03
Cycles (2)−0.02 ± 0.01−0.31 ± 0.050.10 ± 0.03−0.16 ± 0.03
Enzyme conc. (2)NS 0.19 ± 0.090.12 ± 0.10−0.14 ± 0.060.15 ± 0.03
DNA conc. (2)−0.190 ± 0.08−0.076 ± 0.04
Time (2)0.16 ± 0.040.14 ± 0.05−0.062 ± 0.02−0.039 ± 0.01
Temp. × cycles0.09 ± 0.04−0.11 ± 0.07−0.10 ± 0.08NS 0.047 ± 0.03
Temp. × enzyme  conc.NS NS NS −0.072 ± 0.03
Temp. × DNA conc.NS NS
Temp. × time−0.11 ± 0.06NS −0.12 ± 0.02−0.12 ± 0.02
Cycles × enxyme  conc.0.16 ± 0.100.090 ± 0.05
Cycles × DNA conc.−0.08 ± 0.04
Cycles × time0.089 ± 0.05
Enzyme conc. × timeNS −0.14 ± 0.05NS

[i] Regression coefficients are given with the confidence limits atP = 0.05. (nt) = Nucleotides.