Team:Pitt/Skin Probiotic/Dehydrogenase/Methods

Methods

1. Double restriction digestion with the restriction enzymes EcoRI, SpeI, XbaI, or PstI
   We prepared the double restriction digestion according to table 1. After thoroughly yet gently pipetting the mixture up and down several times, the reaction was then incubated at 37 ℃ for 30 minutes. After incubated, the reaction was immediately loaded onto a pre-made 0.8% agarose gel.
   Table 1: Reagent Table for Double Restriction Digestion.
   

   Components	Amount (ul)
   Sterilized Double Distilled Water (SDDW)	13
   Fast Digest Buffer	2
   Plasmid DNA	7
   Enzyme 1	1
   Enzyme 2	1
   Total	23

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