Gene analysis of multiple oral bacteria by PCR-capillary polymer
electrophoresis
Chenchen Liu1,2, Yoshinori Yamaguchi2,3, Shinichi Sekine4,5, Zhenqing Li1, Yi Ni2,
Xifang Zhu6, Xiaoming Dou2,6
1
Engineering Research Center of Optical Instrument and System, University of
Shanghai for Science and Technology, 516 Jungong Road, Shanghai 200093, China.
2
Institute of Photonics and Bio-medicine (IPBM), Graduate School of Science, East
China University of Science and Technology (ECUST), 130 Meilong Road, Shanghai
200237, China.
3
Department of Applied Physics, Graduate School of Engineering, Osaka University,
Yamadaoka, Suita-city, Osaka, 565-0871, Japan.
4
Division of Special Care Dentistry, Osaka University Dental Hospital, Osaka
University, 1-8 Yamadaoka, Suita, Osaka, 565-0871, Japan.
5
Department of Preventive Dentistry, Osaka University Graduate School of Dentistry
Osaka University, 1-8 Yamadaoka, Suita, Osaka, 565-0871, Japan.
6
School of Optoelectronic Engineering, ChangZhou Institute of Technology, 299
South Tongjiang Road, Changzhou, Jiangsu 213002, China.
Correspondence: Dr. Yoshinori Yamaguchi, Department of Applied Physics, Graduate
School of Engineering, Osaka University, Yamadaoka, Suita-city, Osaka, 565-0871,
Japan. Tel: (06)6879-7837
Dr. Xiaoming Dou, Graduate School of Science, East China University of Science and
Technology, 130 Meilong Road, Shanghai 200237, China.
Email: yoshi.yamaguchi@ap.eng.osaka-u.ac.jp; xiaomingdou@yeah.net.
SI. 1.
Table S 1. Primers and lengths for target genes of P.g, T.d and T.f.
Gene target of
bacteria
Porphyromonas
gingivalis
Sequence 5' ---------3'
PCR product
(bp)
Fw
GTAGATGACTGATGGTGAAAACC
Rv ACGTCATCCCCACCTTCCTC
197
Tannerella
forsythia
Fw GCGTATGTAACCTGCCCGCA
Rv TGCTTCAGTGTCAGTTATACCT
641
Treponema
denticola
Fw AAGGCGGTAGAGCCGCTCA
Rv AGCCGCTGTCGAAAAGCCCA
311
SI. 2. CPE was processed by 5 runs to evaluate the precision performance. Internal
standard was 500 bp DNA fragment.
Figure S1. Electropherogram and fitting curve of 100 bp DNA ladder marker. CPE conditions
were the same as in Fig. 2.
SI. 3. Raw data for linear regression test of CPE.
Figure S2. Electropherogram of periodontal pathogens PCR products in different diluted
concentration (A) 15%, (B) 10%, (C) 8%, (D) 6%, (E) 4%, (F) 2%, (G) 0%. The injected
periodontal pathogens PCR products mixture contained 100 bp DNA ladder marker at 6.5 ng μL-1
concentration and TBE at 0.5x concentration. The injection time was 5 s. Other CPE conditions
were the same as in Fig. 1.
SI. 4.
Table S2. Calculated data for quantifying P.g (197 bp) T.d (311 bp) and T.f (641 bp),
the internal standard was 500 bp DNA fragments with 1.5 ng μL-1 concentration.
Target gene size (bp)
197/200
311
641
AreaTimes/(AreasTime)
1.04
0.27 0.49
Integration standard error (E-04)
2.76
2.45 3.90
Concentration (ng μL-1)
1.56
0.40 0.73