Kais Khudhair al Hadrawi
Tris base: A buffering agent used to maintain a stable pH. Tris base is usually added at a concentration of around 50-100 mM. Calcium chloride (CaCl2): Calcium ions are often included in MCLB-1 buffer to stabilize nucleic acids and proteins. The concentration of CaCl2 is typically in the range of 5-10 mM. Magnesium chloride (MgCl2): Magnesium ions are essential cofactors for many enzymatic reactions, including those involved in DNA and RNA processing. MgCl2 is usually added at a concentration of 1-5 mM. Lithium chloride (LiCl): Lithium ions can be included in MCLB-1 buffer to enhance the precipitation of nucleic acids. The concentration of LiCl is typically around 50-100 mM. Bromophenol blue: This dye is often added to MCLB-1 buffer as a tracking dye for agarose gel electrophoresis. The concentration of bromophenol blue is usually very low, around 0.01%. Water: The remaining volume of the buffer is made up of distilled water or nuclease-free water to achieve the desired final volume and concentration of the buffer components.
The MCLB-1 buffer, also known as Modified Chelat Buffer, is commonly used in molecular biology and biochemistry for various applications such as protein extraction, enzyme assays, and DNA isolation. The composition of MCLB-1 buffer typically includes the following ingredients:
Here's a general recipe for preparing MCLB-1 buffer:
- Tris base: 50-100 mM
- Calcium chloride (CaCl2): 5-10 mM
- Magnesium chloride (MgCl2): 1-5 mM
- Lithium chloride (LiCl): 50-100 mM
- Bromophenol blue: 0.01%
- Water: To make up the final volume
To prepare the buffer, dissolve the appropriate amounts of each component in distilled water, adjust the pH if necessary (typically to around pH 7.5), and sterilize the solution by filtration or autoclaving if needed. Store the buffer at the appropriate temperature and protect it from contamination
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