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Fifteen microliters of serially diluted stock extracts were transferred correspondingly into watery plates containing 225 μl of sterile H2O.
Briefly, the stock extracts (100 mg/ml) were prepared in nutrient broth and serially diluted as 50 mg/ml, 25 mg/ml, 12.5 mg/ml and 6.25 mg/ml.
Prior to the day of test, the stock plates were prepared by serially diluting (2 fold dilution) the stock extracts (5 mg/ml) to 7 point concentration (ranging from 5 to 0.16 mg/ml) with DMSO or sterile H2O (for H2O extracts only).
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Kelp stock extracted slowly and carefully from costly Rishiri kombu, fabulous sea salt (her preferred source is from Vietnam), perfectly brewed domestic rice vinegar and artisanal miso and soy sauce enable the chef to draw out the innate flavors of the plant world.
To establish a highly sensitive protocol for effective huNoV detection in waters, 16 published and newly designed reverse transcription polymerase chain reaction (RT-PCR) primer pairs specific for huNoV genogroup I (GI) and genogroup II (GII) were comparatively evaluated side-by-side using single sources of huNoV RNA stock extracted from local clinical isolates.
Stock extract (600 mg/ml) was used in this assay.
The concentration of the stock extract was determined as required.
They were then impregnated with 10 μl of various stock extract solution.
The stock extract for MBS was prepared by redissolving in dimethyl sulfoxide (DMSO) 0.1%%, (BIO BASIC INC., NY, USA).
The stock extract for MBS was prepared by redissolving the MBS extract powder in dimethyl sulfoxide (DMSO) 0.1%, (BIO BASIC INC., NY, USA).
Prior to the bioassay, working solutions of the extracts were prepared by diluting stock extract solutions to four times the maximum desired final testing concentration in sterile 7H9 broth.
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