Selected article for: "expression vector and final concentration"
Author: Yu, Xiaobo; Song, Lusheng; Petritis, Brianne; Bian, Xiaofang; Wang, Haoyu; Viloria, Jennifer; Park, Jin; Bui, Hoang; Li, Han; Wang, Jie; Liu, Lei; Yang, Liuhui; Duan, Hu; McMurray, David N.; Achkar, Jacqueline M.; Magee, Mitch; Qiu, Ji; LaBaer, Joshua
Title: Multiplexed Nucleic Acid Programmable Protein Arrays
  • Document date: 2017_9_20
    • ID: 7t1o19kn_16
    Snippet: All human and viral ORF plasmids were obtained from DNASU (https://dnasu.org/), and transferred into a T7-based mammalian expression vector, pANT7-cGST, as previously described [18, 30, 33] . Purified DNA plasmids were prepared by our automated DNA factory robot as previously described [18, 30, 33] , and were normalized to 1,200 ng/μL, such that multiplexed plasmids contributed equally to the final concentration. In other words, a plasmid in a f.....
    Document: All human and viral ORF plasmids were obtained from DNASU (https://dnasu.org/), and transferred into a T7-based mammalian expression vector, pANT7-cGST, as previously described [18, 30, 33] . Purified DNA plasmids were prepared by our automated DNA factory robot as previously described [18, 30, 33] , and were normalized to 1,200 ng/μL, such that multiplexed plasmids contributed equally to the final concentration. In other words, a plasmid in a five-multiplexed spot would represent 240 ng/μL. Five (5) different plasmids containing a different gene-of-interest were mixed with a master printing mixture containing BSA (Sigma), BS 3 cross-linker (Thermo Fisher Scientific, IL) and polyclonal α-GST antibody (Thermo Fisher Scientific, IL) [26] , and subsequently incubated at 4 o C for 2 h. M-NAPPA and NAPPA were printed by the NAPPA Protein Array Core (http://nappaproteinarray.org/) according to published protocols [18, 30, 33] . The quality of printed plasmid DNA on M-NAPPA and NAPPA was determined using PicoGreen DNA staining [26] .

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