Recipe
- Add 1ml of 0.1% Diethylpyrocarbonate (DEPC) to 1000ml distilled water.
- Mix well and let set at room temperature for 1 hour.
- Autoclave.
- Let cool to room temperature prior to use.
What is DEPC water?
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. Diethyl pyrocarbonate.
How do you use DEPC treated water?
DEPC treatment of water usually involves adding 0.2 ml DEPC to every 100 ml of water, shaking vigorously to dissolve (the DEPC falls to the bottom), and then autoclaving to drive off the remaining DEPC. This “DEPC-treated” water can then be used as RNAse-free water.
Can I use DEPC treated water for PCR?
IBI DEPC Treated Water is ideal for all applications in a molecular biology lab including PCR, RT-PCR, restriction enzyme assays, modifying enzyme assays, transfection, cloning, transformation and all general molecular biology lab procedures.
How do you make DEPC water? – Related Questions
When should you not use DEPC water?
Do not add DEPC to any buffer containing mercaptans or 1° amine groups, with which DEPC is reactive (Berger, 1975). Perhaps the most common buffers to which DEPC exposure is to be avoided are the Tris (tris(hydroxymethyl)aminomethane) buffers.
How do you make 0.1 DEPC water?
Add 100 µl of DEPC to 100 ml dH2O for a final concentration of 0.1% DEPC in a glass container. 2. Mix well and let set at room temperature for 1 hour.
Can I use DEPC treated water for DNA?
Description: DEPC-treated Water is highly pure and nuclease-free (tested for both contaminating nonspecific DNase and RNase activities), specially developed for all RNA work.
Is DEPC water PCR grade?
DEPC-treated water is not recommended for PCR or in vivo experiments. (For PCR applications, Teknova’s molecular biology–grade water for PCR is tested for RNase and DNase activity.)
Why is DEPC treated water not recommended for PCR?
It does sound like contamination–remember that DEPC only inactivates RNases that are present in water when its made, and won’t inactivate RNases that are introduced after its been autoclaved. Residual DEPC that hasn’t been inactivated after autoclaving can also inhibit RT and PCR reactions (or so I’ve heard).
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.
Is DEPC water deionized?
DEPC(Diethylpyrocarbonate)-treated deionized water for RNase-free applications. DEPC inactivates RNase enzymes in water by covalent modifications of the histidine residues.
What is the difference between DEPC water and nuclease free water?
Posted May 9, 2020. Nuclease-free water is free of both DNAse and RNAse, and involves treating with DEPC (diethylpyrocarbonate) and/or autoclaving to inactivate RNAse and DNAse. Ultrapure water, on the other hand, is obtained by ultrafiltration to achieve the high level of purity. It also does not contain any nucleases
Why deionized water is used in PCR?
Deionized water used in PCR is usually sterilized by autoclaving for 15 minutes at 121∘C and 15 psi [5]. After autoclaving, heat-stable DNA from dead cells would contaminate subsequent processes such as PCR mixtures and primer aliquots.
Can I use distilled water instead of deionized water?
No. Deionised water and distilled water are two of the most common types of treated water. Both have very high purity levels and can be used for similar applications, so while many assume they’re the same thing, this isn’t the case. Deionised water and distilled water are both created in different ways.
Is boiled water deionised?
Deionized (DI) water is water that has been treated to remove all ions – typically, that means all of the dissolved mineral salts. Distilled water has been boiled so that it evaporates and then re-condensed, leaving most impurities behind. Distillation is one of the oldest methods for creating pure water.
Is distilled water same as deionized water?
Distilled water has been boiled to create purified water free of contaminants and impurities like chlorine and dissolved solids. Deionized (DI) water, like distilled water, is also very pure. Where they differ is that deionized water is water that has had all the dissolved mineral particles removed from it.
Is boiled water the same as distilled water?
No, they aren’t the same. Boiled water is simply water that has had the temperature increased until it reaches its boiling point. This kills a host of bacteria that can make a person sick. Distilled water is water that has been stripped of all impurities, including minerals and microorganisms.
Can you drink DI water?
Yes, you can drink distilled water. However, you might not like the taste because it’s flatter and less flavorful than tap and bottled waters. Companies produce distilled water by boiling water and then condensing the collected steam back into a liquid. This process removes impurities and minerals from the water.
What pH is deionized water?
pH of High Quality Deionized Water
Very high quality deionized water may produce a pH measurement of 6.0 to 6.4 following this test procedure.
Can deionized water grow bacteria?
Growth of microorganisms from deionized water on R2A agar (Figure 2) revealed that a large variety of microbes were present in deionized water and were viable.