Supplemental On-Line Information for Developing a Qpcr Method to Quantify Ahr-PCP-DNA Complexes

Supplemental On-line Information for “Developing a qPCR method to quantify AhR-PCP-DNA complex for detection of environmental trace-level PCP” submitted to Ecotoxicology

Authors: Xiaoxiang Zhao*, Xiaoqian Pang, Nuanapa Chaisuwan

Affiliation of all authors: College of Environmental Science and Engineering, Donghua University, 201620 Shanghai, People’s Republic of China.

Table S1 DNA sequences of the PCR primers used in this study

Sequence（5’-3’） / Products
F1:CGG AGT TCC GTG AGA AGA GGA TAA CGC AGG AAA GAA CAT G / 924 bp
F2: CTC TTC TCA CGC AAC TCC GGT CAG GCA ACT ATG GAT GAA C
R1:TCG ACG CTC AAG TCA GAG / 359 bp
R2:TAG CAC CGC CTA CAT ACC

Table S2 Components of receptor-ligand-DNA complex

Sample / Ligand / Ah Receptor / DNA Probe
2 / 10 µL PCP / 10 µL / 2 µL
3 / 10 µL Glycerol / 10 µL / 2 µL
4 / 10 µL DMSO / 10 µL / 2 µL
5 / 10 µL PCP / 10 µL H2O / 2 µL
6 / 10 µL H2O / 10 µL / 2 µL

Figure S1 SDS PAGE of cytosol and Ah receptors obtained from Hepatic cytosol. From left to right: protein marker, hepatic cytosol receptors and purified Ah receptors

Figure S2 DNA probes by PCR amplification. Lanes (from left to right) represent DNA markers and

924 bp DNA probes

Figure S3 Representative set of melting curves obtained by plotting fluorescence data against their cycle numbers

Figure S4 1.5% Agarose gel electrophoreses by PCR amplification for sample 2-6 in components of receptor-ligand-DNA complex. Lane 1 is DNA ladder marker (2 kb, 1 kb, 750 bp, 500 bp, 250 bp and 100 bp)

Figure S5 Amplification curve of fluorescence qPCR detection lines from left to right represented 10-9 to 10-13 M of PCP (A) and Standard curve of PCP (B)

Figure S5A

Figure S5B

Figure S6 Characteristic wavelength of 10-4 M PCP