Transgenic rice TT51-1 transformation event foreign vector integration site complete sequence and use thereof
A technology for transgenic rice and transformation events, applied in the field of full sequence of exogenous vector integration sites of transgenic rice TT51-1 transformation events, can solve problems that have not yet been discovered
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[0046] 1. PCR amplification of the full sequence of the exogenous vector integration site in the transgenic rice TT51-1 transformation event
[0047] First preheat 20% SDS to 65°C, take 15ml SDS extraction buffer (0.1 TrisHCl, 0.05MEDTA, 1M NaCl pH8.0) and add it to a 50ml centrifuge tube, then add 2.5μl β-mercaptoethanol and mix well; liquid nitrogen Grind about 3g of leaves, transfer the powder to 50ml of a 50ml centrifuge tube containing extraction buffer, shake and mix on a shaker, add 2ml of preheated 20% SDS, mix well, and bathe in 65°C water for at least 30 minutes, during which time Shake the test tube gently; after the water bath, quickly place the centrifuge tube on ice, add 3ml 3M KAc, mix well, and place it on ice for 30 minutes; centrifuge at 5000g at 4°C for 5min; transfer the supernatant to a new 50ml centrifuge tube, Add 2 / 3 volume of isopropanol, mix well, place at -20°C for more than 30min; centrifuge at 6000g, 4°C for 15min, pour off the supernatant, wash wi...
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