DEPC


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DEPC

(organic chemistry)
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After washing the papers with 70% ethanol, the miRNAs were eluted in DEPC water at 95 [degrees]C for 30 min, and subjected to RT-qPCR.
DNA stock solutions (100 [micro]M) were prepared in DEPC deionized water and stored in small aliquots.
The reaction system contained 12.5 [micro]L of Bio-Rad iQ SYBR Green Supermix, 1 [micro]L of sense primer (50 pg), 1 [micro]L of antisense primer (5pg), 1 [micro]L of cDNA, and 9.5 [micro]L of DEPC [H.sub.2]O, resulting in a total volume of 25 [micro]L.
Briefly, 10 [micro]l of each PCR product was mixed with 2 [micro]l of Tango buffer, 1 [micro]l of restriction enzyme, and 12 [micro]l of DEPC water.
Due to the high initial DNA concentration obtained in the Genomic DNA extraction kits Mini Kit and Wizard[R] Genomic DNA Purification Kit, there was need for a serial dilution of [10.sup.-1], [10.sup.-2], [10.sup.-3] in sterile DEPC water.
The supernatant was discarded and the RNA was washed in 1 mL of 75% ethanol diluted in diethylpyrocarbonate (DEPC)-treated water.
In many cases, the water is first chemically treated with diethylpyrocarbonate (DEPC) and then autoclaved to ensure the eradication of nuclease activity.
Amplification was performed with final reaction volume of25l, including Master mix, DEPC, reverse and forwardprimers and DNA sample.Finally, PCR was carried out with the following steps.
Subsequently, the precipitate was dried in vacuum after washing with 75% ethanol, and then dissolved in DEPC water.
Tri Reagent (Sigma, DB) was used to purify mRNA, which was kept in water containing diethyl pyrocarbonate (DEPC).
Pepsin (0.01% in 0.01 M HCl) was used to treat the tissue for 5 minutes at 37[degrees]C, and a brief rinse in DEPC treated [H.sub.2]O followed.
TRIzol was obtained from Invitrogen; diethyl pyrocarbonate (DEPC) was acquired from AppliChem.