A kind of Daqu bacteria dgge standard band making method
A production method, bacteria technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems that harmful microorganisms cannot be suppressed, and the repeatability of microorganisms cannot be strictly guaranteed, so as to avoid the waste of time and funds.
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Embodiment 1
[0040] Embodiment 1 Preparation of Daqu DGGE standard band
[0041] 1. Obtain the bacterial DGGE map during the fermentation process of Daqu, and determine the dominant bacteria in the fermentation process of Daqu.
[0042] l) Select qualified samples from different fermentation stages of Daqu.
[0043] During the fermentation process of Daqu, the mother koji, the raw koji, the first turned over Daqu, the second turned over Daqu and the outbound koji were selected as research samples.
[0044]2) Extract total microbial DNA.
[0045] The extraction method is as follows: Weigh 0.4g of mother koji, raw koji, the first overturned Daqu, the second overturned Daqu and out of warehouse koji samples, respectively add about 200mg of glass beads, and use the OMEGA soil DNA extraction kit to extract Total DNA was extracted from five samples. Add 1000ul SLX solution, vortex at high speed for 5min to mix evenly, add 100ul DS solution, and vortex to mix. Water bath at 70°C for 10 minute...
Embodiment ２
[0057] Example 2 Identification of microbial populations of Daqu DGGE standard bands during Daqu fermentation
[0058] 1) In the qualitative detection of microbial populations in the fermentation stage, 5 samples of mother koji, raw koji, first-time overturned koji, second-time overturned koji, and koji out of warehouse were taken as samples to be tested. The 16S rDNA of the samples to be tested The PCR amplification of the V3 variable region uses nested PCR amplification, and the specific method is the same as above. The PCR reaction conditions for the amplification of the DGGE standard bands were as follows: using the DGGE bands corresponding to the selected common dominant bacteria as templates, pre-denaturation at 94°C for 4 min, then denaturation at 94°C for 45s, primer annealing at 50-55°C for 45s, 72°C Primers were extended for 1min30s at ℃, cycled 30 times, and finally extended at 72℃ for 10min; PCR products were detected by 1% agarose gel electrophoresis, and the elec...
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