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文献和实验Alkaline Southern Blotting Procedure
. When ready to load, add an appropriate amount of 10X tracking dye, no EtBr. Pour the gel: Mix 2 grams ultra pure agarose in 250 ml 1X TAE (.8%). Heat in microwave to boiling. Continue boiling and swirling until agarose is completely dissolved. Cool gel
Western Blotting with Alkaline Phosp
NaOH. 1.For transfer of proteins smaller than 20 kDa, transfer proteins from gel to PVDF (polyvinylidene difluoride) membrane at 1Amp constant current for 45 mins or equivalent (250mAmp for 3 hours or 500mAmp for 90 minutes) in transfer buffer (25mM
10.3 , add concentrated urea and concentrated NaCl separately to yield a final concentration of 7 M urea and 3 M NaCl. Centrifuge the clear solution at 85,500 xg for 48 hours at 4° C to pellet extracted DNA.
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