gDNA
To isolate genomic DNA (gDNA) Gentaur Delos Labs uses the following Protocol
Sample Collection: Collect the tissue or cells from which you want to isolate genomic DNA. This could be blood, tissue biopsy, cultured cells, plant material, etc. Ensure proper handling and storage of the samples to maintain DNA integrity.
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Cell Lysis: Disrupt the cell membranes to release the genomic DNA. This can be achieved through various methods depending on the sample type, such as:
- Mechanical disruption (grinding, homogenization)
- Chemical lysis (using detergents)
- Enzymatic lysis (using proteases or lysozyme)
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Removal of Proteins and RNA: After cell lysis, remove proteins and RNA from the lysate using proteinase K treatment and RNase digestion, respectively. This step ensures that only DNA remains in the sample.
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DNA Purification: Purify the genomic DNA from the lysate to remove contaminants and impurities. Commonly used methods include:
- Phenol-chloroform extraction: Organic solvents are used to separate DNA from proteins and other cellular components.
- Silica membrane-based spin column purification: DNA selectively binds to the silica membrane while contaminants are washed away.
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Washing and Elution: Wash the DNA-bound membrane or beads to remove residual impurities and then elute the purified genomic DNA using an appropriate buffer solution.
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Quantification and Quality Assessment: Measure the concentration and assess the quality of the isolated genomic DNA using methods such as spectrophotometry (e.g., UV absorbance at 260 nm) or fluorometry (e.g., using DNA-specific dyes). Verify the purity and integrity of the DNA by checking the A260/A280 ratio and running an agarose gel electrophoresis, respectively.
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Storage: Store the purified genomic DNA at an appropriate temperature (-20°C or -80°C) to maintain its stability and integrity for future experiments.
Comparison of Eleven Methods for Genomic DNA Extraction Suitable for Large-Scale Whole-Genome Genotyping and Long-Term DNA Banking Using Blood Samples
Gentaur supplies the protocols and reagents used for genomic DNA isolation with optimized protocols and reagents for efficient DNA isolation from different sample types.