Tris(hydroxymethyl)aminomethane → Sulfuric acid, monododecyl ester, potassium salt (1:1)
Reaction equation
C(O)C(N)(CO)CO.O=C[C@@H]([C@H]([C@@H]([C@@H](CO)O)O)O)O.[OH-].[Na+].[S:23]([O-:39])([O:26][CH2:27][CH2:28][CH2:29][CH2:30][CH2:31][CH2:32][CH2:33][CH2:34][CH2:35][CH2:36][CH2:37][CH3:38])(=[O:25])=[O:24].[Na+].C([O-])(=O)C.[K+:45]>>[S:23]([O-:39])([O:26][CH2:27][CH2:28][CH2:29][CH2:30][CH2:31][CH2:32][CH2:33][CH2:34][CH2:35][CH2:36][CH2:37][CH3:38])(=[O:24])=[O:25].[K+:45]Participants
Procedure
The cells are thawed, where appropriate, and then lysed. Chemical lysis can be broken down into three steps. The first consists in resuspending the cells in a 25 mM Tris, pH 6.8, 50 mM glucose, 10 mM ETDA buffer or equivalent. The cells are then lysed in a mixture containing 0.2 M NaOH and 1% SDS. The pH of the solution is approximately 12. The choice of an ionic detergent is essential since a non-ionic detergent gives extraction yields which are 10 times lower. The lysis is followed by a pseudo-neutralization of the medium in the presence of potassium acetate (the final pH of the solution is between 5.5 and 6). This acidification of the medium results in the appearance of a precipitate containing the proteins and part of the chromosomal DNA and the RNA. This precipitation is due to reaction of the sodium dodecyl sulphate (SDS) with the potassium acetate, which together form a white precipitate of potassium dodecyl sulphate.
Workup
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Sources & evidence
- ORD record
- ord-d0f6b2ac822347d0b0ca30a510833f60
- Source dataset
- uspto-grants-2004_05
- Patent number
- US06730781B1
Reaction notes
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