By the time you use DEPC treated reagents, the chemical has already degraded into CO2, ethanol, and water. If it's not present, then RNases - being the resilient enzymes they are - can spontaneously renature. What am I missing?
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#2
hobglobin
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Posted 09 November 2012 - 01:04 PM
But the RNAse is "killed" by DEPC, by modifying -NH, -SH, and -OH groups in the proteins. After treatment you usually autoclave the solution and get rid of the remaining DEPC (autoclaving breaks down the molecule) because it should not be there later when you work with RNA ("traces of DEPC modify purine residues (A+G) in RNA by carboxymethylation"; Openwetware).
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Ahrenhase
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Posted 10 November 2012 - 12:01 AM
hobglobin, on 09 November 2012 - 01:04 PM, said:
But the RNAse is "killed" by DEPC, by modifying -NH, -SH, and -OH groups in the proteins. After treatment you usually autoclave the solution and get rid of the remaining DEPC (autoclaving breaks down the molecule) because it should not be there later when you work with RNA ("traces of DEPC modify purine residues (A+G) in RNA by carboxymethylation"; Openwetware).
Ah, chemical modification, got it. Thanks.
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ayman hesham
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Posted 11 January 2013 - 04:57 AM
thank you but we should not use any buffer containing amines like TRIS -EDTA because these buffer will interfere with action of DEPC water
