What does DEPC water do in PCR?
DEPC-treated Water – DEPC destroys enzymatic activity by modifying -NH2, -SH and -OH groups in RNases and other proteins. DEPC treatment is a very effective way to treat solutions that will contact RNA.
How is water treated with DEPC?
DEPC Treated Water Recipe
- Add 1ml of 0.1% Diethylpyrocarbonate (DEPC) to 1000ml distilled water.
- Mix well and let set at room temperature for 1 hour.
- Let cool to room temperature prior to use.
What kind of water do you use for PCR?
PCR-grade Water is intended for use in molecular biology applications including PCR and RT-PCR. The ultra-pure and sterile filtered water is manufactured free of detectable inhibitors, contaminants or enzymatic activity.
What does DEPC stand for?
|DEPC||Depth Control Log (energy production)|
|DEPC||Data Evaluation and Publication Committee|
|DEPC||Department of Environmental Pollution Control (various nations)|
|DEPC||Department for Environment, Planning and the Countryside (UK)|
How do you treat DEPC tips?
It is not necessary to treat your tips and tubes with DEPC, as long as sterilization at about 200 ℃. Add 0.1% DEPC in ddH2O. Mix it with magnetic strirer for about an hour in dark. Dip all the tips and tubes completely merge in 0.1% DEPC water overnight in dark.
What is DEPC water used for PCR?
Thermo Scientific deionized, diethylpyrocarbonate (DEPC) treated and 0.22 µm membrane-filtered water. It is ideal for applications involving RNA. • Free of endo-, exodeoxyribonucleases, ribonucleases, and phosphatases. • DEPC-treated Water is not recommended for PCR or in vivo experiments.
Is the RT-PCR grade water ready to use?
The RT-PCR Grade Water is ready to use and requires no preparation, mixing, or autoclaving. RT-PCR Grade Water is tested for prokaryotic as well as eukaryotic genomic DNA contamination using an ultrasensitive PCR assay.
What are the limitations of DEPC treated water?
• Free of endo-, exodeoxyribonucleases, ribonucleases, and phosphatases. • DEPC-treated Water is not recommended for PCR or in vivo experiments. For Research Use Only. Not for use in diagnostic procedures.
Does DEPC treatment affect the sensitivity of PCR?
DEPC treatment would be preferred for enhance sensitivity and completely remove any potential for RNA degradation by RNases during cDNA synthesis, but nuclease free water should be sufficient for PCR. However, DEPC treatment will not affect your PCR, if you choose to use this, as long as the DEPC has been inactivated.