A kind of anaerobic river channel sediment degrading bacteria and its application
A technology of river sediment and strains, applied in the field of environmental microorganisms, can solve problems such as human health and ecological environment risks, and achieve the effects of reduction and harmlessness, convenient operation, and low cost
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Embodiment 1
[0030] Example 1: Isolation, screening and performance measurement of anaerobic river sediment degrading bacteria
[0031] The media and components used are as follows:
[0032] Liquid medium: glucose 5 g / L, tryptone 3 g / L, beef meal 2 g / L, NaCl 4 g / L, K 2 HPO 4 0.75 g / L, KH 2 PO 4 0.75 g / L, FeSO 4 ·7H 2 O 0.05 g / L, MgSO 4 ·7H 2 O 0.3 g / L, (NH 4 ) 2 SO 4 3 g / L, cysteine 0.25 g / L, natural pH.
[0033] Solid medium: glucose 5 g / L, tryptone 3 g / L, beef meal 2 g / L, NaCl 4 g / L, K 2 HPO 4 0.75 g / L, KH 2 PO 4 0.75 g / L, FeSO 4 ·7H 2 O 0.05 g / L, MgSO 4 ·7H 2 O 0.3 g / L, (NH 4 ) 2 SO 4 3 g / L, cysteine 0.25 g / L, 15 g / L agar powder, natural pH.
[0034] Channel sediment liquid culture medium: channel sediment (total solid TS content is 11%, VS content is 7.8% of TS, both are mass fractions), add water to adjust the TS content of the sediment to 5%, and the pH value is 7.0.
[0035] Channel sediment solid medium: channel sediment (TS content is 11%, VS content...
Embodiment 2
[0042] Example 2: Biological identification of anaerobic river sediment degrading strain YE6
[0043] The genomic DNA of strain YE6 was extracted and used as a template to amplify the 16S rDNA of the strain using a pair of universal primers (27F, 1492R). The upstream primer was 27F (5'-AGAGTTGATCCTGGCTCA-3') and the downstream primer was 1492R (5'-GGTTACCTTGTTACGACTT-3').
[0044] The PCR reaction system (20 μL) is as follows: template DNA 0.5 μL, PCR Taqmix 10 μL, upstream and downstream primers 0.6 μL each, plus ddH 2 O to the reaction system to 20 μL.
[0045] PCR program: pre-denaturation at 94 °C for 5 min, denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, extension at 72 °C for 1 min 30 s, the above cycle was repeated 30 times, extension at 72 °C for 10 min, and finally stored at 4 °C.
[0046] The PCR product was purified and sequenced by Shanghai Jie Li Biotechnology Co., Ltd., and the 16S rDNA sequence of the obtained strain was shown in SEQ ID NO.1. The...
Embodiment 3
[0048] Example 3: Preparation of liquid bacterial agent for anaerobic river sediment degrading strain YE6
[0049] The pure YE6 strain was inoculated into 10 mL of liquid medium, cultured anaerobic at 37 °C for 24 h, and then transferred to the liquid medium of the next-level expansion culture with an inoculum of 5% of the liquid medium volume. Under the same conditions, multi-stage anaerobic expansion culture can be carried out to obtain the liquid inoculum of the strain.
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