Hydrogen Producing Bacteria Lab Report

Hydrogen Producing Bacteria was incubated in a complete - mix digester with work volume 1.7 L, seeded with sludge obtained from the local sewage treatment plant. Each liter of feed medium was composed of the following : 7 g of glucose, 1 g NaHCO3 , 500 mg of NH4Cl , 250 mg KH2PO4 , 250 mg K2HPO4 , 320 mg of MgSO4 • 7H2O , 50 mg of FeCl 3 , NiSO4 32 mg , 50 mg CaCl2, Na2BO7 7.2 mg H2O , 14.4 mg (NH4) 6MO7O24 H2O , 23 mg of ZnCl2 , 21 mg CoCl2 H2O , 10 mg CuCl2•2H2O and 30 mg of MnCl2•4H2O . The reaction medium is maintained at 4° C, purged with nitrogen to ensure anaerobic conditions , and continuously fed by means of a peristaltic pump. The fermenter was maintained at 36° C in the dark to inhibit photosynthetic activity. The mixed liquid was
The injector, column and detector temperatures were maintained at 57° C, 180° C and 50° C, resp. The concentrations of volatile fatty acids and alcohols were determined by a second GC thereof model, which was equipped with a flame ionization detector and a 10 mx 0.53 mm Hewlett Packard FFAP fused silica capillary column, according to the procedure described by Yu and Fang (2000) procedures. The glucose concentration was measured by the phenol- sulfuric acid acid method (Herbert et al. 1971). The volatile suspended solids (VSS) were measured according to standard methods (APHA
Among them, the primer sets GM3F / GM4R and EUB968F / UNIV1392R were used in PCR for bacteria while the primer set ARCH622F / ARCH934R was PCR applied for archaea. All PCR amplifications were carried out in 30 pi of a pH 8.3 buffer (Pharmacia Biotech, Piscataway, NJ) containing 200 pM each of the four deoxynucleotide triphosphates, 15 mM MgCl2, 0.1 pM of each primers, and 1 unit Taq polymerase (Pharmacia Biotech, Piscataway, NJ). An automated thermal cycler (GeneAmp PCR 9700, Perkin-Elmer, Foster City, Calif.) Was used for PCR-amplification using the following program denaturing gradient gel electrophoresis (DGGE),