Phenol-Chloroform RNA extraction, Schweisguth Protocol

Published on Author Anne Petzold

Protocol

"Extraction of Drosophila total RNA" @ http://francois.schweisguth.free.fr/protocols/Extraction_Drosophila_total_RNA.pdf

(extraction based on combination of Trizol and Phenol-Chloroform, spinning steps @ 4 DEG)

Samples

app. 12 male fly bodies and app. 30 heads, dissection on CO2 pad @ RT.

Process

Samples frozen @ - 80 DEG in PBS before extraction. Protocol as instructed, but used centrifuge at RT. No DNase I treatment.

Results

Body sample: [260/280] 1.7, 4.25 ug/ul

Brain sample: [260/280] 1.86, 4.27 ug/ul

Indicative of contamination with phenol or protein. Probable sources of contamination: #1 pestle, other: spinning at RT, no DNAse treatment.