Mechanism of microwave sterilization materials and methods
A microwave oven
    QX microwave , model QX-30HM7, rated microwave frequency 2450MHz, rated input power 30KW, maximum output power of 800W.
2 dialysis bag
    3500 MWCO dialysis bag , Item MD34.3.5, flatten when dry width ( half circumference ) 24mm, purchased from Beijing Branch Hongda Biotechnology Co., Ltd. .
Three tested strains
    Black variant of Bacillus subtilis ATCC9372, purchased from the Institute of Microbiology Microbial Culture Collection .
4 medium
Liquid ordinary nutrient agar medium : 109 peptone , sodium chloride 59 , beef powder 159 , distilled water 1000ml.
Solid ordinary nutrient agar medium : 109 peptone , sodium chloride 59 , beef powder, 159 , 209 agar , distilled water 1000ml.
5 Test Preparation of bacterial suspension
    Nutritional cell preparation : 250ml beaker containing 50ml of nutrient broth inoculated Bacillus subtilis 2ml black
ATCC9372 variant spores suspension cultured overnight , the overnight culture was centrifuged at 5000rpm 10min, after centrifugation the precipitate was washed four times with PBS buffer , the pellet was washed with PBS buffer , the pellet was suspended , diluted to the required concentration , into the flask sterilized , dispensed into 7ml centrifuge tube in the back.
    Preparation of Bacillus : Bacillus subtilis the activated nutrient agar medium inoculated with a sterile smooth evenly curved rods , and incubated at 37 ℃ . 5 days incubation at room temperature , the rate of 90 % or more spores when the spores were scraped with a sterile spoon . With sterile cotton wool was filtered to remove the scraped agar . At 45 . C in a shaker shock for 24 hours, to chain scission , dispersed into individual spores of Bacillus subtilis . The Bacillus subtilis centrifuged at 7000rpm 15min, the supernatant decanted , resuspended with distilled water , then centrifuged , repeated twice . In the 8012 water bath for 10min, killing propagules . Then centrifuged at 7000rpm conditions . Spores was diluted with distilled water to the desired concentration , aliquots scraper centrifuge tube 7ml alternate a-15 "C condition F saved. Spores prepared monthly .
    6 bags of dialysis treatment
    The dialysis bag cut into appropriate degrees K (10em) - ^ volume in 2% (m / v) sodium bicarbonate and IrrtmoULEDTA (PH = 8 0) will be boiled dialysis bag IO minutes, rinsed thoroughly with distilled water dialysis state , the dialysis bag set ImfnolfL EDTA (PH = 80) was boiled for 10 minutes, cooled special dialysis bag , stored D 4 "0, J butterfly commercial insurance dialysis bag always immersed in the liquid ( from step dialysis bag from the film to wear gloves when given Yi operation ) , in the State of yan State of distilled water from the mouth will be cleaned outside the dialysis bag .