What is RNase free water?
RNase-Free Water is pure, quality-tested water suitable for use in all experiments that require RNase-free water, including PCR, RT-PCR, and real-time PCR, and is included in most QIAGEN PCR and RT-PCR Kits, such as the Taq PCR Master Mix Kit, the HotStarTaq Plus Master Mix Kit, and the HotStar HiFidelity Polymerase …
Is DEPC-treated water the same as RNase free water?
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 diethylpyrocarbonate for at least 2 hours at 37 °C and then autoclaved (at least 15 min) to inactivate traces of DEPC.
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. 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.
How is RNase free water prepared?
The use of ultrafiltration together with autoclaving allows for the preparation of “RNase-free” water. This process is an effective alternative to DEPC treatment that delivers high purity RNase-free water.. RNases that can be present in water or reagents used during performing molecular biology experiments degrade RNA.
Why is RNase-Free Water used?
The presence of nucleases such as DNase and RNase in water can degrade precious molecular samples and even ruin experiments. To prevent DNA and RNA sample loss, it is essential that highly pure, nuclease-free water be used in applications such as PCR, cDNA synthesis, nucleic acid purification, sequencing, and cloning.
Is DEPC treated water deionized?
Thermo Scientific deionized, diethylpyrocarbonate (DEPC) treated and 0.22 µm membrane-filtered water. It is ideal for applications involving RNA.
Can I use DEPC treated water for Qpcr?
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.
How do you use DEPC water?
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 is DEPC and how it works?
Diethyl pyrocarbonate (DEPC) is an efficient, nonspecific inhibitor of RNases. It is typically used to treat water and solutions before working with easily degraded RNA. DEPC reacts with amine, hydroxy and thiol groups of proteins thereby inactivating RNAses (and other enzymes).
Is DEPC water endotoxin free?
DEPC-Treated Water for those who prefer chemically (DEPC) treated water for PCR applications is also available. Endotoxin Free Water is free of endotoxins and enzymes, including proteases. Certified tested by the Limulus amebocyte lysate (LAL) test for endotoxins and determined to be <0.0050EU/ml..
How do you DEPC water?
How do you use DEPC?
If you have ever worked with RNA, you know about DEPC (diethylpyrocarbonate). You add it to water at a concentration of 0.1%, shake or stir, incubate at 37°C for two hours or at room temperature overnight and, as if targeted by a magic bullet, the RNAses that may have been in the water are gone.
Is there a way to make RNase free from DEPC?
The most cautious approach for making RNase-free solutions would be to mix molecular biology grade powdered reagents up in DEPC-treated water. Alternatively, many pre-made nuclease-free solutions can be purchased from Thermo Fisher Scientific and other companies.
How is DEPC treated water used for RNA?
DEPC-Treated Water is suitable for use with RNA. It is prepared by incubating 0.1% DEPC and is then autoclaved to remove the DEPC. Although autoclaving does inactivate DEPC by causing hydrolysis of DEPC, EtOH (ethanol) and CO 2 are, however, released as reaction by-products.
Which is better 1% DEPC or 0.1% RNase?
TRUE: 1% DEPC is better at removing RNase than 0.1% It is certainly true but higher DEPC concentrations mean that there is likely to be more left-over DEPC/DEPC by-products after autoclaving, and these can inhibit downstream applications.
How does DEPC interact with Tris and RNase?
The solution will still smell of ethanol, but also sweet-smelling volatile esters formed by the reaction of ethanol with trace carboxylic acids. The amino group on tris interacts with DEPC, making it unable to do it’s job of decontaminating the solution. FALSE: 0.1% DEPC is sufficient to inhibit any amount of RNase in solution.