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delivery. LEAPChem is happy to highlight Diethyl pyrocarbonate.
Basic
Information of Diethyl pyrocarbonate
Chemical Name: Diethyl pyrocarbonate
Cas No.: 1609-47-8
Molecular Formula: C6H10O5
Chemical Structure:
Diethyl pyrocarbonate
(DEPC), also called diethyl dicarbonate (IUPAC name), is used in the laboratory
to inactivate RNase enzymes in water and on laboratory utensils. It does so by
the covalent modification of histidine (most strongly), lysine, cysteine, and
tyrosine residues.
DEPC-treated (and therefore RNase-free)
water is used in handling of RNA in the laboratory to reduce the risk of RNA
being degraded by RNases.
Water is usually treated with 0.1% v/v DEPC
for at least 2 hours at 37 °C and then autoclaved (at least 15 min) to
inactivate traces of DEPC. Inactivation of DEPC in this manner yields CO2 and
ethanol. Higher concentrations of DEPC are capable of deactivating larger
amounts of RNase, but remaining traces or byproducts may inhibit further
biochemical reactions such as in vitro translation. Furthermore, chemical
modification of RNA such as carboxymethylation is possible when traces of DEPC
or its byproducts are present, resulting in impaired recovery of intact RNA
even after buffer exchange (after precipitation).
DEPC is unstable in water and susceptible
to hydrolysis to carbon dioxide and ethanol, especially in the presence of a
nucleophile. For this reason, DEPC cannot be used with Tris or HEPES buffers.
In contrast, it can be used with phosphate-buffered saline or MOPS. A handy
rule is that enzymes or chemicals which have active -O:, -N: or -S: cannot be
treated with DEPC to become RNase-free, as DEPC reacts with these species.
Furthermore, DEPC degradation products can inhibit in vitro transcription.
DEPC derivatization of histidines is also
used to study the importance of histidyl residues in enzymes. Modification of
histidine by DEPC results in carbethoxylated derivates at the N-omega-2
nitrogen of the imidazole ring. DEPC modification of histidines can be reversed
by treatment with 0.5 M hydroxylamine at neutral pH.
DEPC can also be used for probing the
structure of double-stranded DNA.
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References:
https://en.wikipedia.org/wiki/Diethyl_pyrocarbonate
https://pubchem.ncbi.nlm.nih.gov/compound/diethyl_pyrocarbonate
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC339840/
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