| Autor: |
Bhoomi, Madhu, Mohammed Farhan, Lakdawala, Tina L, Gumienny |
| Rok vydání: |
2022 |
| Předmět: |
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| Zdroj: |
Journal of visualized experiments : JoVE. (184) |
| ISSN: |
1940-087X |
| Popis: |
Genomic DNA extraction from single or a few Caenorhabditis elegans has many downstream applications, including PCR for genotyping lines, cloning, and sequencing. The traditional proteinase K-based methods for genomic DNA extraction from C. elegans take several hours. Commercial extraction kits that effectively break open the C. elegans cuticle and extract genomic DNA are limited. An easy, faster (~15 min), and cost-efficient method of extracting C. elegans genomic DNA that works well for classroom and research applications is reported here. This DNA extraction method is optimized to use single or a few late-larval (L4) or adult nematodes as starting material for obtaining a reliable template to perform PCR. The results indicate that the DNA quality is suitable for amplifying gene targets of different sizes by PCR, permitting genotyping of single or a few animals even at dilutions to one-fiftieth of the genomic DNA from a single adult per reaction. The reported protocols can be reliably used to quickly produce DNA template from a single or a small sample of C. elegans for PCR-based applications. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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