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Volume 27 Issue 6
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Article Navigation >  Journal of Pharmaceutical Practice and Service  > 2009  >  27(6): 421-425

YANG Juan, GUO Mei-Li. Choice of restriction endonuclease and establishment of silver-staining method in cDNA-AFLP System for Carthamus tinctorius[J]. Journal of Pharmaceutical Practice and Service, 2009, 27(6): 421-425.
Citation: YANG Juan, GUO Mei-Li. Choice of restriction endonuclease and establishment of silver-staining method in cDNA-AFLP System for Carthamus tinctorius[J]. Journal of Pharmaceutical Practice and Service, 2009, 27(6): 421-425.
shu

Choice of restriction endonuclease and establishment of silver-staining method in cDNA-AFLP System for Carthamus tinctorius

  • Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China

  • Received Date: 2009-06-18
  • Objective :To establishing an appropriate combination of restriction endonuclease and silver-staining system for cDNA-AFLP analysis in Carthamus tinctorius L,. Methods :The efficiency of combination of PstI/MseI and MseI/EcoRI is compared.Several key factors in silver-staining including the affiliation and dissection dosage in sequencing plate processing,TEMED dosage in gel,pre-electrophoresis,temperature factor,staining liquid configuration etc are researched and discussed. Results :Enzyme efficiency of PstI/MseI is higher than MseI/EcoRI.Sequencing plate should be cleaned thoroughly.TEMED volume should be moderate.30mins pre-electrophoresis should be carried out.Temperature should be constant through the whole electrophoresis process. Conclusion :This research has found a high effective restriction endonuclease combination can further discover the genetic background of safflower and has established an appropriate silver-staining system which has overcome several problems in staining process.
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Choice of restriction endonuclease and establishment of silver-staining method in cDNA-AFLP System for Carthamus tinctorius

  • Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China
  • Received Date: 2009-06-18

Abstract: Objective :To establishing an appropriate combination of restriction endonuclease and silver-staining system for cDNA-AFLP analysis in Carthamus tinctorius L,. Methods :The efficiency of combination of PstI/MseI and MseI/EcoRI is compared.Several key factors in silver-staining including the affiliation and dissection dosage in sequencing plate processing,TEMED dosage in gel,pre-electrophoresis,temperature factor,staining liquid configuration etc are researched and discussed. Results :Enzyme efficiency of PstI/MseI is higher than MseI/EcoRI.Sequencing plate should be cleaned thoroughly.TEMED volume should be moderate.30mins pre-electrophoresis should be carried out.Temperature should be constant through the whole electrophoresis process. Conclusion :This research has found a high effective restriction endonuclease combination can further discover the genetic background of safflower and has established an appropriate silver-staining system which has overcome several problems in staining process.

YANG Juan, GUO Mei-Li. Choice of restriction endonuclease and establishment of silver-staining method in cDNA-AFLP System for Carthamus tinctorius[J]. Journal of Pharmaceutical Practice and Service, 2009, 27(6): 421-425.
Citation: YANG Juan, GUO Mei-Li. Choice of restriction endonuclease and establishment of silver-staining method in cDNA-AFLP System for Carthamus tinctorius[J]. Journal of Pharmaceutical Practice and Service, 2009, 27(6): 421-425.
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