Identification of Genes Involved in Flocculation by Whole Genome Sequencing of Thauera aminoaromatica Strain MZ1T Floc-defective Mutants Page: V
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2.6 Research O bjectives.........................................................................................................35
CHAPTER 3 M ATERIALS AND M ETHODS ...........................................................................................37
3.1 Bacterial Plasm ids and Strains..........................................................................................37
3.2 Culture conditions and storage........................................................................................40
3.3 M edia and Chem icals.................................................................................................... 40
3.4 DNA M anipulation Techniques........................................................................................43
3.5 Next Generation Sequencing...........................................................................................43
3.5.1 Ion Library and em plate preparation..............................................................43
3.5.2 Ion sequencing...................................................................................................44
3.6 Bioinform atics Analysis.....................................................................................................44
3.7 SNP Validation by Sanger Sequencing............................................................................45
3.8 M utant Com plem entation................................................................................................45
3.9 EPS Purification..................................................................................................................47
3.10 Q uantification of EPS......................................................................................................48
3.11 FTIR Analysis.....................................................................................................................48
3.12 Deacetylation of MZ1T EPS............................................................................................48
3.12 Gene Expression Analysis...............................................................................................49
3.12.1 RNA Extraction and cDNA Synthesis.............................................................49
3.12.2 Droplet Digital PCR..........................................................................................50
CHAPTER 4 RESULTS.............................................................................................................................53
4.1 Floc-Defective Mutants MZ1T 39A and 20A Genome Sequencing............................53
4.2 SNP Validation Using PCR and Sanger Sequencing.......................................................68V
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Prombutara, Pinidphon. Identification of Genes Involved in Flocculation by Whole Genome Sequencing of Thauera aminoaromatica Strain MZ1T Floc-defective Mutants, dissertation, December 2015; Denton, Texas. (https://digital.library.unt.edu/ark:/67531/metadc822777/m1/6/?rotate=180: accessed July 17, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; .