Preparation method of nano-gold, nano-gold and application
A nano-gold, reaction technology, applied in the field of bioengineering, to achieve the effect of wide adaptability and moderate size
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Embodiment 1
[0034] Bacterial culture and preparation of nano gold synthesis agent.
[0035] (1) Bacterial culture: Deinococcus radiodurans with the preservation number ATCC13939 was plated and activated for culture, then a single colony was picked and inoculated in 5 mL of TGY medium, and cultivated overnight at 32°C on a shaker with a shaker speed of 220rpm then transferred to a large bottle of medium (1L culture bottle containing 500mL TGY medium) in a ratio of 1:100, and cultivated for 24h at 32°C and 220rpm to make the bacteria culture to the stationary phase (OD600nm=1.0).
[0036] (2) Collecting and washing the bacteria: After the bacteria are cultivated to a stable period, they are collected by centrifugation at 8000 g for 10 minutes to obtain fresh bacteria. It was resuspended in 0.01M phosphate buffer solution with a pH of 7.0, fully shaken, centrifuged at 8000 g for 10 min, and the cells were collected.
[0037] (3) Disintegrating the cells and preparing cell extracts: resuspen...
Embodiment 2
[0040] Bacterial culture and preparation of nano gold synthesis agent.
[0041] (1) Bacterial culture: Deinococcus radiodurans with the preservation number ATCC13939 was plated and activated for culture, then a single colony was picked and inoculated in 5 mL of TGY medium, and cultivated overnight at 30°C on a shaker with a shaker speed of 220rpm Then transfer to large bottle medium (1L culture bottle containing 500mL TGY medium) in the ratio of 1: 100, cultivate 24h under the condition of 30 ℃, 220rpm and make bacterial culture to stationary phase (OD600nm=1.0).
[0042] (2) Collecting and washing the bacteria: After the bacteria are cultivated to a stable period, they are collected by centrifugation at 8000 g for 10 minutes to obtain fresh bacteria. It was resuspended in 0.01M phosphate buffer solution with a pH of 7.0, fully shaken, centrifuged at 8000 g for 10 min, and the cells were collected.
[0043](3) Disintegrating the cells and preparing cell extracts: resuspend th...
Embodiment 3
[0046] Preparation of gold nanoparticles.
[0047] (1) Preparation of synthetic reaction solution: 1mM Au 3+ Solution (auric chloride, HAuCl 4 ·3H 2 O) In a 50mL centrifuge tube, add an appropriate amount of Deinococcus radiodurans extract dry powder (prepared in Example 1, without ammonium sulfate treatment) so that the concentration is 3.00mg / mL, at room temperature (pH 7, 25°C) Make it fully mixed and reacted under the conditions.
[0048] (2) Monitoring the reaction process: As the reaction proceeds, the color of the solution changes, gradually becomes purple and then tends to be stable. After 6 hours of reaction, the wavelength scanning is performed (there is a peak at 545nm).
[0049] (3) Purification of nano-gold solution: filter the above-mentioned purple solution with a 0.22 μm syringe filter, centrifuge at 15000 g for 30 min, collect the precipitate, wash with distilled water several times, and dialyze for 24 h with a dialysis bag with a cutoff of 14 KD to remove ...
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