Table 1 Summary and expected behavior of the ten tested protocols according to extraction buffer and polymerization type (chemical or photopolymerization) used.
Extraction Buffer | Extraction temperature | Expected behaviour | Protocol name | Conditions | |
|---|---|---|---|---|---|
Chemical Polymerization (Temed + Ammonium Persulfate) | Photopolymerization (Methylene blue) | ||||
SDS/Tris (pH 7.5) | 70 °C | Reference protocol [Muller et al., 2016] | STC | STP | Basic SDS |
SDS/Glycine (pH 2.5) | 70 °C | Remove DNA (at pH 2.5 DNA is less negatively charged and precipitate in contact with SDS) | SGC | SGP | Acidic SDS |
Urea/Spermine/CHAPS (pH 8) | RT | Remove DNA (Precipitate in contact with Spermine) | USC | USP | Chaotropic agent |
CTAC/Glycine (pH 2.5) | 70 °C | Remove interfering anionic macromolecules by precipitation | / | CTP | Cationic detergent |
CTAC/Glycine/Urea 4 M (pH 2.5) | RT | Remove interfering anionic macromolecules by precipitation | / | CUP | |
HEPES/KCl/EDTA/Spermidine/SB3–14 | 0 °C | Weak denaturation + Remove nucleus/DNA | NAC | NAP | Native |
