Denaturing Gel-Loading Buffer without Dye [4X]
Cat. No. DCP-DGLBWOD4XStorage: -20°C (Recommend to keep it cold at all time)
Size: 5 X 1ml (500 loadings: 10ul Loading Buffer + 30ul sample = total loading volume 40ul / well)
Appearance: clear, colorless liquid
Sterile: DEPC treated Ultrapure Water Type 1 used; Filtered 0.22 micron membrane once.
Denaturing Gel-Loading Buffer has formamide as the key component. Formamide is a strong denaturing agent helps denature secondary structures in nucleic acids. It is useful for working with ssDNA, DNA and RNA with expected secondary structure when running agarose gel electrophoresis. Formamide is also an ionizing solvent and an RNA stabilizer in gel electrophoresis. For example, the Denaturing Gel-Loading Buffer enables denaturing of the nicked plasmid harboring the DNA of interest. This product contains EDTA which is a chelating agent that binds divalent metal ions like Ca2+ and Mg2+, inhibiting metal-dependent enzymatic reactions and protecting nucleic acids. EDTA is used to prevent degradation of DNA and RNA and to inactivate nucleases.
This Denatureing Gel Loading Buffer is used to increase the density of your sample solution which enables the sample solution to sink into the wells while maintaining the denatured form of ssDNA, DNA or RNA samples. For ssDNA works, DNAs derived from the nicked plasmid keep single-strand status. The secondary structure must be denatured before it enters the denaturiong gel of choice. The Denaturing Gel-Loading Buffers are specifically designed for use with denaturing agarose or polyacrylamide gel electrophoresis (PAGE) of DNA and RNA samples. The formamide denatures the nucleic acids, allowing them to migrate based on size rather than secondary structure.
Non-denaturing agarose gel electrophoresis can analyze RNA integrity and overall presence/absence without denaturing the samples. Denaturing electrophoresis with a formamide-based loading buffer is the preferred method for most applications requiring resolving individual RNA species. The buffer is typically added to the nucleic acid sample and mixed, and the whole sample load is heated to 95°C to complete the denaturation process. Once the sample cools (in some cases while hot), the sample is loaded onto the denaturing agarose oro polyacrylamide gel for electrophoresis.
* This product DOES NOT include any component of dyes (i.e., SDS, bromophenol blue, and xylene cyanol) for tracking the migration of samples.
Quality Control:
- Sterility: Filtered 0.22 micron membrane once.
- DNase activity: None DNase activity was detected after incubation plasmid DNA and this product for 18 hr at room temperature.
- RNase activity: DEPC-treated Ultrapure Water Type1 used; None RNase activity was detected after incubation ribosomal RNA and this product for 18 hr at room temperature.
Storage and Shelf Life:
- Storage: -20 degree Celsius
- Shelf life: 3 years
All DiagnoCine Precision Sterile media are filtered-sterilized with a 0.1-micron filtration two times, plus 0.04 micron once. Thus, mycoplasma contamination is prevented. The smallest size mycoplasma type can be about 0.2 microns.
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