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文献和实验Fastfilter Plasmid Maxi Kit Spin Protocol
to completely neutralize the solution. Complete neutralization of the solution is vital of obtaining good yields. 6. Add 8.3 ml Buffer GBT, cover, and gently mix by inverting tube 3-5 times. 7. Prepare the HiBind Maxi Column. Place a HiBind Maxi
Preparation of Affinity Column
ml on nylon (0.22uM poresize),using Buchner funnel and vacuum.Do not allow beads to dry. 5.Wash bed with 3x2ml dH2O. 6.Scrape 0.5-1ml into 15ml microfuge tube. 7.Add 2.22ml CREBtide ligand solution to tube. Rock at 4℃ for 4hrs. 8.Transfer slurry to column
Preparation of Affinity Column制备亲和层析柱【UCSF】
1.Add 222.2μl 1M MOPS,pH 7.5 to 2ml of 10mg of CREBtide (0.1M MOPS pH 7.5 final concentration). 2.Read OD205,OD280 (using 0.1M MOPS buffer as blank) 3.Affigel-10 is stored frozen at 4.Filter 1-2ml on nylon (0.22uM poresize),using Buchner funnel
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