RIKEN BioResource Center Gene Engineering Division RIKEN DNA Bank catalog # Microbial Genomic DNA T Strain: JCM 7501 Organism: Thermoanaerobacter thermocopriae JGD Lot # : 12533 12533_A3K8 Date : 2015/11/19 1. Shipping Volume: 30 ul 2. Concentration of Genomic DNA 45.7 1.9 Concentration (OD260): A260/A280: ng/ul 3. Electrophoresis of Genomic DNA Electrophoresis : 1% agarose gel, 1x TAE buffer Loading DNA amount : 4, 2, 1 ul/lane Size marker : Lambda/HindIII (200 ng/lane) 2 1 Marker Lambda/HindIII (NEB#N3012S) 0.5 (ul) 23.1 6.6 9.4 4.4 2.0 2.3 (Kb) 0.6 4. PCR Amplification 2015/11/19 Date : Conditions of PCR ribosomal RNA (rRNA) gene Template : 100 fold diluted DNA, 1 ul Enzyme : KOD Plus (Toyobo) Volume : 20 ul Reaction program: 94 94 x30 cycle 60 68 68 degC 2 min degC 15 sec degC 30 sec degC 60 sec degC 2 min PCR primers 16S rRNA Eubacteria set EB-20F 5' AGTTTGATCCTGGCTC 3' EB-350F 5' TACGGGAGGCAGCAG 3' EB-1100R 5' AGGGTTGCGCTCGTTG 3' EB-1400R 5' ACGGGCGGTGTGTAC 3' EB-1530R 5' AAGGAGGTGATCCAGCC 3' (1) (2) (3) (4) (5) (6) Sample Number EB-1100R EB-1400R EB-1530R EB-20F (1) (2) (3) EB-350F (4) (5) (6) Electrophoresis : 1% agarose gel, 1x TAE buffer Loading DNA amount : 3 ul PCR product/lane Size marker : Lambda/HindIII (200 ng/lane) 5. Nucleotide sequence of rRNA gene □Confirmed date : 2015/11/27 http://dna.brc.riken.jp
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