Haynes Lab:Notebook/Engineering PC-TFs/2015/05/07

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Summary Miniprep'd liquid cultures. All samples between 100-250 ng/μL. Digest and Gel Purification Reagent Volume DNA (2μg) 23μL 12μL 12.0 μL 12.0 μL 20.0 μL 16.0 μL 20.0 μL (From well 2 to well 8) 10X buffer 3.0 μL 3.0 μL 3.0 μL 3.0 μL 3.0 μL 3.0 μL 3.0 μL XbaI 0μL 0μL 0μL 0μL 2μL 2μL 2μL PstI 2μL 2μL 2μL 2μL 2.0μL 2μL 2μL SpeI 2μL 2μL 2μL 2μL 0.0 μL 0μL 0μL dH2O 0μL 11μL 11μL 11μL 3μL 7μL 7μL Total 30 μL --> 37°C/ 15 min. Ladder 15 μL/lane; 1% agarose; Expected: All linkers (KAH021-024) [SpeI/PstI] V0120 + linker (very short). Only one band expected. Inserts (xPcD-RFP-VP64; KAH0879,KAH089,KAH090 [XbaI/PstI] 1053bp Observed lengths: W1: Ladder W2: KAH021 W3: KAH022 W4: KAH023 W5: KAH024 W6: KAH087 W7: KAH089 W8: KAH090