AAV Helper-Free Virus Purification

According to a previously published protocol (28 (link)), we packaged the AAV helper-free system (Agilent Technologies, Santa Clara, CA), the pRep-Cap (AAV5; Applied Viromics, Fremont, CA), and the pHelper plasmid. AAV was purified using a cesium chloride density gradient, combined with ultracentrifugation. Virus titer was determined using quantitative real-time polymerase chain reaction analysis (SYBR green; Takara Bio Inc., Shiga, Japan).

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Obi-Nagata K., Suzuki N., Miyake R., MacDonald M.L., Fish K.N., Ozawa K., Nagahama K., Okimura T., Tanaka S., Kano M., Fukazawa Y., Sweet R.A, & Hayashi-Takagi A. (2023). Distorted neurocomputation by a small number of extra-large spines in psychiatric disorders. Science Advances, 9(23), eade5973.

Publication 2023

AAV helper-free system SYBR green

Cesium chloride Plasmid Quantitative real time polymerase chain reaction analysis Sybr green Ultracentrifugation

Corresponding Organization : Gunma University

Other organizations : University of Pittsburgh, The University of Tokyo, Showa University, Sophia University, University of Fukui

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Variable analysis

independent variables

AAV helper-free system (Agilent Technologies, Santa Clara, CA)
PRep-Cap (AAV5; Applied Viromics, Fremont, CA)
PHelper plasmid

dependent variables

Virus titer

control variables

Cesium chloride density gradient
Ultracentrifugation

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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