Selection and use of melatonin supporting bacteria to reduce infantile colic
A melatonin, baby technology, applied in bacteria, medical raw materials derived from bacteria, enzymes, etc., can solve problems such as frequent recommendation
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[0186] Method for identifying strains with genes encoding cell surface localized 5'-nucleotidases
[0187] Bacteria were grown on MRS plates for 16 h at 37 °C in an anaerobic atmosphere. Ten bacterial colonies were collected with a sterile plastic loop and suspended in 100 microliters of sterile water (PCR quality). (Alternative: prepare DNA from bacterial culture using a suitable method).
[0188] The presence of the 5'-nucleotidase gene can be checked by PCR, for example by using PuReTaq Ready ToGo PCR beads (GE HealthCare) and either of the primer pairs described below (0.4 mM each). The bacterial suspension or DNA preparation (0.5 µl) should be added to the PCR mix and the PCR reaction should be performed by running the following program: 95 °C, 5 min; 30x (95 °C, 30 sec; 55 °C, 30 sec; 72°C, 30 seconds); 72°C, 10 minutes. PCR products can be separated and visualized by using standard agarose gel electrophoresis and sequenced by standard Sanger sequencing (using the for...
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