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CE of Small DNA Fragments Using Linear Polyacrylamide Matrices

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Abstract

Mutations at codons 248 and 249 of p53 gene showed a relatively high incidence in gastric cancer patients. Development of novel methods for the detection of codon mutations is of great importance for gastric cancer research. Studies have showed that the separation matrix can significantly influence the separation efficiency and resolution of small DNA fragments in CE. In order to achieve baseline separation of PCR-amplified products of small DNA fragments from gastric cancer tissue, linear polyacrylamides (LPA I and LPAII) were designed and synthesized in the current study. LPAI and LPAII were used as separation matrixes to separate small size fragments (less than 70 bp) of pBR322/BsuRI DNA Marker and the separation conditions were optimized. Optimum separations were performed at 25 kV in reversed-polarity mode with capillary temperature set at 15 °C. The signal of DNA fragments was detected using laser-induced fluorescence detector, with an argon ion laser as the excitation source that emits at 488 nm. A 520 nm bandpass filter was used as an emission cut-off filter. The resolution of small DNA fragments was higher when LPAI was used as separation matrix compared to LPAII, accompanied with longer migration time. The results indicated that LPAI as separation matrix was more efficient for the separation of small DNA fragments (less than 70 bp) than other LPAs. A rapid and sensitive analysis method for the separation and detection of small DNA fragments (less than 70 bp) was established in this study. The method was successfully applied to detect the mutations at codons 248 and 249 in p53 gene from gastric cancer tissues.

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References

  1. Ren JC (2000) J Chromatogr B Analyt Technol Biomed Life Sci 741:115–128. doi:10.1016/S0378-4347(00)00090-6

    Article  CAS  Google Scholar 

  2. Loeb LA (1996) Mutat Res 350:279–286. doi:10.1016/0027-5107(95)00117-4

    Google Scholar 

  3. Sokurenko EV, Tchesnokova V, Yeung AT, Oleykowski CA, Trintchina E, Hughes KT, Rashid RA, Brint JM, Moseley SL, Lory S (2001) Nucleic Acids Res 29:e111. doi:10.1093/nar/29.22.e111

    Article  CAS  Google Scholar 

  4. Zhong WW, Yeung ES (2002) J Chromatogr A 960:229–239. doi:10.1016/S0021-9673(01)01393-0

    Article  CAS  Google Scholar 

  5. Wimmer K, Zhu XX, Lamb BJ, Kuick R, Ambros P, Kovar H, Thoraval D, Elkahloun A, Meltzer P, Hanash SM (2001) Med Pediatr Oncol 36:75–79. doi:10.1002/1096-911X(20010101)36:1<75::AID-MPO1018>3.0.CO;2-O

    Article  CAS  Google Scholar 

  6. Nachamkin I, Panaro NJ, Li M, Ung H, Yuen PK, Kricka LJ, Wilding P (2001) J Clin Microbiol 39:754–757. doi:10.1128/JCM.39.2.754-757.2001

    Article  CAS  Google Scholar 

  7. Hu CT, O’Shaughnessy KM (2001) Electrophoresis 22:1063–1068. doi:10.1002/1522-2683()22:6<1063::AID-ELPS1063>3.0.CO;2-S

    Article  CAS  Google Scholar 

  8. Kuklin A, Munson K, Gjerde D, Haefele R, Taylor P (1997) Genet Test 1:201–206

    Article  CAS  Google Scholar 

  9. Arakawa H, Nakashiro S, Maeda M, Tsuji A (1996) J Chromatogr A 722:359–368. doi:10.1016/0021-9673(95)00430-0

    Article  CAS  Google Scholar 

  10. Wenz HM (1994) Nucleic Acids Res 22:4002–4008. doi:10.1093/nar/22.19.4002

    Article  CAS  Google Scholar 

  11. Wulfert M, Tapprich C, Gattermann N (2006) J Chromatogr B Analyt Technol Biomed Life Sci 831(1–2):236–247. doi:10.1016/j.jchromb.2005.12.024

    CAS  Google Scholar 

  12. Sivakumaran TA, Kucheria K, Oefner PJ (2003) Curr Sci 84:291–296

    CAS  Google Scholar 

  13. Han FT, Huynh BH, Ma YF, Lin BC (1999) Anal Chem 71:2385–2389. doi:10.1021/ac990160x

    Article  CAS  Google Scholar 

  14. Wenz HM, Dailey D, Johnson MD (2001) Methods Mol Biol 163:3–17

    CAS  Google Scholar 

  15. Marsh M, Tu O, Dolnik V, Roach D, Solomon N, Bechtol K, Smietana P, Wang L, Li X, Cartwright P, Marks A, Barker D, Harris D, Bashkin J (1997) J Capillary Electrophor 4:83–89

    CAS  Google Scholar 

  16. Lindberg P, Roeraade J (2001) Methods Mol Biol 163:289–308

    CAS  Google Scholar 

  17. Khrapko K, Coller HA, Li-Sucholeiki XC, Andre PC, Thilly WG (2001) Methods Mol Biol 163:57–72

    CAS  Google Scholar 

  18. Dovichi NJ, Zhang J (2001) Methods Mol Biol 167:225–239

    CAS  Google Scholar 

  19. Cheng J, Mitchelson KR (1994) Anal Chem 66:4210–4214. doi:10.1021/ac00095a016

    Article  CAS  Google Scholar 

  20. Dolnik V (1999) J Biochem Biophys Methods 41:103–119. doi:10.1016/S0165-022X(99)00041-X

    Article  CAS  Google Scholar 

  21. Zhu MD, Hansen DL, Burd S, Gannon F (1989) J Chromatogr A 480:311–319. doi:10.1016/S0021-9673(01)84300-4

    Article  CAS  Google Scholar 

  22. Sartori A, Barbier V, Viovy JL (2003) Electrophoresis 24:421–440. doi:10.1002/elps.200390052

    Article  CAS  Google Scholar 

  23. Kourkine IV, Hestekin CN, Buchholz BA, Barron AE (2002) Anal Chem 74:2565–2572. doi:10.1021/ac020025b

    Article  CAS  Google Scholar 

  24. Jia H, Jia ZP, Fu MQ, Wang R, Chen QY, Xie H, Ma J (2006) J Lanzhou Univ 32:7–11

    CAS  Google Scholar 

  25. Barron AE, Soane DS, Blanch HW (1993) J Chromatogr A 652:3–16. doi:10.1016/0021-9673(93)80639-P

    Article  CAS  Google Scholar 

  26. Song LG, Liang DH, Fang DF, Chu B (2001) Electrophoresis 22:1987–1996. doi:10.1002/1522-2683(200106)22:10<1987::AID-ELPS1987>3.0.CO;2-K

    Article  CAS  Google Scholar 

  27. Heller C (1999) Electrophoresis 20:1978–1986. doi:10.1002/(SICI)1522-2683(19990701)20:10<1978::AID-ELPS1978>3.0.CO;2-G

    Article  CAS  Google Scholar 

  28. Tseng WL, Chang HT (2001) Electrophoresis 22:763–770. doi:10.1002/1522-2683(200102)22:4<763::AID-ELPS763>3.0.CO;2-W

    Article  CAS  Google Scholar 

  29. Gao QF, Yeung ES (2000) Anal Chem 72:2499–2506. doi:10.1021/ac991362w

    Article  CAS  Google Scholar 

  30. Slater GW, Guillouzic S, Gauthier MG, Mercier JF, Kenward M, McCormick LC, Tessie FR (2002) Electrophoresis 23:3791–3816. doi:10.1002/elps.200290002

    Article  CAS  Google Scholar 

  31. Chen QY, Wang R, Jia ZP, Liu YY, Xie H, Ma J, Wang YQ (2007) Chinese. J Anal Chem 35:1305–1308

    Google Scholar 

  32. Chu B, Ying QC, Wang Y, Zhang J (2005) Macromol Sympos 227:77–88. doi:10.1002/masy.200550907

    Article  CAS  Google Scholar 

  33. Heller C (2001) Electrophoresis 22:629–643. doi:10.1002/1522-2683(200102)22:4<629::AID-ELPS629>3.0.CO;2-S

    Article  CAS  Google Scholar 

  34. Ozawa S, Sugano K, Sonehara T, Fukuzono S, Ichikawa A, Fukayama N, Taylor M, Miyahara Y, Irie T (2004) Anal Chem 76:6122–6129. doi:10.1021/ac049385k

    Article  CAS  Google Scholar 

  35. Du M, Flanagan JH Jr, Lin BC, Ma YF (2003) Electrophoresis 24:3147–3153. doi:10.1002/elps.200305572

    Article  CAS  Google Scholar 

  36. Xu F, Jabasini M, Baba Y (2002) Electrophoresis 23:3608–3614. doi:10.1002/1522-2683(200210)23:20<3608::AID-ELPS3608>3.0.CO;2-3

    Article  CAS  Google Scholar 

  37. Zhou P, Yu SB, Liu ZH, Hu JM, Deng YZ (2005) J Chromatogr A 1083:173–178. doi:10.1016/j.chroma.2005.05.096

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported by the foundation from National Natural Science Foundation of China (No.20775089) and National Natural Science Foundation of Gansu province.

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Authors and Affiliations

  1. Department of Pharmacy, Lanzhou General Hospital of PLA, Lanzhou, Gansu, China

    Zheng-Ping Jia, Rong Wang, Qiao-Yun Chen, Hua Xie, Jun Ma & Juan Wang

  2. Jiamusi College, Heilongjiang University of Traditional Chinese Medicine, Jiamusi, China

    Qiao-Yun Chen

  3. Life Sciences College, Lanzhou University, Lanzhou, China

    Qiao-Yun Chen & Yuan-Yuan Liu

Authors
  1. Zheng-Ping Jia
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  2. Rong Wang
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  4. Hua Xie
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  7. Juan Wang
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Correspondence to Rong Wang.

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Jia, ZP., Wang, R., Chen, QY. et al. CE of Small DNA Fragments Using Linear Polyacrylamide Matrices. Chroma 70, 1127–1134 (2009). https://doi.org/10.1365/s10337-009-1280-z

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