Amplifying the human NANOG promoter

The human NANOG promoter was amplified from pNanog-Luc (Addgene #25900) using the forward primer hNan-F2 (or -F1) and the reverse primer hNan-R1 (or -R2); the primers in parentheses are fluorescently labeled versions of the listed primers; see Supplementary Table 2. The 404 bp PCR fragments were purified using QIAquik Gel Extraction Kit (Qiagen). The pGL-NanogP-5E minus plasmid^{53 (link)} containing the mouse 1535 bp NANOG promoter and 1337 bp enhancer (−5 kbp from NANOG promoter) was digested with PvuI (NEB) to linearize the plasmid. The DNA fragment containing NANOG promoter and enhancer was purified using QIAquik Gel Extraction Kit (Qiagen).

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Sharma R., Choi K.J., Quan M.D., Sharma S., Sankaran B., Park H., LaGrone A., Kim J.J., MacKenzie K.R., Ferreon A.C., Kim C, & Ferreon J.C. (2021). Liquid condensation of reprogramming factor KLF4 with DNA provides a mechanism for chromatin organization. Nature Communications, 12, 5579.

Publication 2021

pNanog-Luc QIAquik Gel Extraction Kit

Human Mouse Plasmid Primers

Corresponding Organization : Baylor College of Medicine

Other organizations : Lawrence Berkeley National Laboratory

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Variable analysis

independent variables

Amplification of the human NANOG promoter from pNanog-Luc (Addgene #25900) using forward primer hNan-F2 (or -F1) and reverse primer hNan-R1 (or -R2)
Digestion of the pGL-NanogP-5E minus plasmid containing the mouse 1535 bp NANOG promoter and 1337 bp enhancer with PvuI (NEB) to linearize the plasmid

dependent variables

None explicitly mentioned

control variables

None explicitly mentioned

controls

Positive control: None mentioned
Negative control: None mentioned

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