Total RNA Extraction from Frozen Brain Tissue

Total RNA was extracted from frozen brain tissues as described previously (18 (link)). Initially, approximately 20 to 30 mg of tissue was snap-frozen in liquid nitrogen and disintegrated using a cryoPREP impactor (Covaris, UK). The pulverized tissue was solubilized in preheated lysis buffer AL and RNA was extracted using the RNAdvance tissue kit (Beckman Coulter, Germany) in combination with a KingFisher Flex purification system (Thermo Fisher Scientific, Germany).

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Pfaff F., Breithaupt A., Rubbenstroth D., Nippert S., Baumbach C., Gerst S., Langner C., Wylezich C., Ebinger A., Höper D., Ulrich R.G, & Beer M. (2022). Revisiting Rustrela Virus: New Cases of Encephalitis and a Solution to the Capsid Enigma. Microbiology Spectrum, 10(2), e00103-22.

Publication 2022

cryoPREP impactor RNAdvance tissue kit KingFisher Flex purification system

Brain Buffer Frozen Nitrogen Tissue

Corresponding Organization : Friedrich-Loeffler-Institut

Other organizations : Lower Saxony State Office for Consumer Protection and Food Safety, German Oceanographic Museum

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Variable analysis

independent variables

Extraction of total RNA from frozen brain tissues

dependent variables

Not explicitly mentioned

control variables

Amount of tissue used (20 to 30 mg)
Snap-freezing of tissue in liquid nitrogen
Disintegration of tissue using a cryoPREP impactor
Solubilization of pulverized tissue in preheated lysis buffer AL
RNA extraction using the RNAdvance tissue kit
Purification using a KingFisher Flex purification system

controls

No positive or negative controls were explicitly mentioned in the provided information.

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