Aux_Aug_14_1_pyro
NGS sequencing of microorganisms present in a high pH spent nuclear fuel pond
Extracted DNA from  high pH pond water samples was amplified for the 16S rRNA gene using primers  27F (5-AGAGTTTGATCCTGGCTCAG-3) (Lane 1991) and 907R  (5-CCGTCAATTCMTTTRAGTTT-3) (Muyzer 1995). Each 50?l PCR reaction mixture  contained 1?l DNA template, primers (0.4?M each), deoxynucleoside  triphosphates (dNTPs; 200?M), 1.5mM MgCl2, 5?l 10X PCR buffer, 5?l bovine  serum albumin (BSA), and 0.5?l Taq polymerase. Template DNA was initially  denatured at 94C for 4 minutes, followed by 30 cycles of denaturing (30  seconds at 94C), annealing (30 seconds at 55C), and extension (60 seconds  at 72C), and a final extension at 72C for 5 minutes. Amplified DNA was  sequenced for Bacterial 16S ribosomal RNA using the Roche 454 Pyrosequencing  platform (Research and Testing Laboratories, Austin, TX, USA).
Aux_Aug_14_1_pyro
SUB5263014
Aux_Aug_14_1_pyro