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Study Of Synthetic Oligodeoxynucleotides Acid By Liquid Chromatography-mass Spectrometry

Posted on:2006-01-20Degree:DoctorType:Dissertation
Country:ChinaCandidate:H Y ChenFull Text:PDF
GTID:1100360215450786Subject:Environmental Science
Abstract/Summary:PDF Full Text Request
Single nucleotide polymorphism (SNP) genotying and deoxyribonucleic acid (DNA) methylation are significant in molecular biology, but their tradition analyse methods have some limitations, and liquid chromatography - mass spectrometry (LC-MS) are favourable for analyzing them. Few reports have focused on studying SNP genotyping and DNA methylation by LC-MS and LC-MS/MS. In this thesis, synthetic oligodeoxynucleotides acid (ODNs) and methylated oligodeoxynucleotides should be analyzed by high performance liquid chromatography - tandem mass spectrometry (HPLC-MS/MS), and the methods of ODNs sequencing and locating methylated sites by mass spectrometry should be built primarily. The studies are exploring and hard, and these can be the base of analyzing DNA adducts by mass spectrometry. The followings are the main content:1. Utilize reversed-phase ion-pair high performance liquid chromatography (RP-IP-HPLC) to separate ODNs, and discuss the retention behavior of ODNs in RP-IP-HPLC from length, nucleotide compositions, variety of 3'-end nucleotide, secondary structure of ODNs, and the different mobile phase system.2. Utilize HPLC-MS/MS to quickly acquire the molecular weight of ODNs, and to ensure the sequence at suitable experimental conditions. Then summarizes the rule of ODNs by MS and MS/MS.3. Utilize HPLC-MS/MS to analyze two groups of ODNs which can be seen as SNP genotypings.4. Utilize HPLC-MS/MS to analyze methylated ODNs.5. Moreover, we have debugged built-in-house time of flight mass spectrometry (TOFMS), and try to analyze ODNs by it.From the five researches above, the following conclusions can be drawn:1. In the RP-IP-HPLC, the shorter and the lower (G+C)% of ODNs are, the earlier they elute from the chromatogram column; At 25℃, the ODNs with different 3'-end nucleotide elute in the order of G
Keywords/Search Tags:high performance liquid chromatography - tandem mass spectrometry (HPLC-MS/MS), Single nucleotide polymorphism (SNP) genotying, deoxyribonucleic acid (DNA) methylation, time of flight mass spectrometry (TOFMS)
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