Accedi

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow specific protocols even for genomic DNA extraction.

Types of genomic DNA extraction methods

The main aim of genomic DNA extraction methods is to separate gDNA from proteins, RNA, and other cell content. It involves four basic steps - 1. Disruption of the cell structure mechanically or using chemicals to obtain the cell lysate 2. Protection of DNA from degradation during processing 3. Separation of the soluble DNA from cell debris 4. Elution of purified DNA.

Most genomic DNA isolation protocols are either solution-based or solid-phase extraction methods. Solution-based methods rely on precipitation and centrifugation steps to separate DNA from other cellular material, followed by organic extraction or "salting out" to separate soluble DNA from cellular proteins. The final DNA precipitation is done using ethanol. In contrast, solid-phase extraction methods use solid support, such as silica or cellulose matrices, to bind DNA, followed by washing and DNA elution from the solid support. It involves centrifugation, vacuum, or magnetic methods to separate the bound DNA from other cellular components.

The choice of gDNA extraction method depends on the type of sample, the number of samples to be processed at once, and the downstream application of the DNA.

Tags

DNA IsolationDNA ExtractionPlasmid DNAGenomic DNALysis MethodLong range DNA SequencingYield Of High quality DNAGenomic DNA Extraction MethodsDisruption Of Cell StructureProtection Of DNASeparation Of Soluble DNAElution Of Purified DNASolution based MethodsSolid phase Extraction MethodsPrecipitation And Centrifugation StepsOrganic ExtractionSalting OutEthanol Precipitation

Dal capitolo 15:

article

Now Playing

15.2 : DNA Isolation

Studio del DNA e dell'RNA

36.9K Visualizzazioni

article

15.1 : DNA recombinante

Studio del DNA e dell'RNA

16.5K Visualizzazioni

article

15.3 : Elettroforesi del DNA su gel di agarosio

Studio del DNA e dell'RNA

91.9K Visualizzazioni

article

15.4 : Marcatura delle sonde di DNA

Studio del DNA e dell'RNA

8.0K Visualizzazioni

article

15.5 : Southern Blot

Studio del DNA e dell'RNA

17.6K Visualizzazioni

article

15.6 : Microarray di DNA

Studio del DNA e dell'RNA

17.0K Visualizzazioni

article

15.7 : DNA complementare

Studio del DNA e dell'RNA

5.5K Visualizzazioni

article

15.8 : Ibridazione fluorescente in situ - FISH

Studio del DNA e dell'RNA

19.1K Visualizzazioni

article

15.9 : PCR - Polymerase Chain Reaction

Studio del DNA e dell'RNA

80.9K Visualizzazioni

article

15.10 : Real Time RT-PCR

Studio del DNA e dell'RNA

56.2K Visualizzazioni

article

15.11 : RACE - Rapid Amplification of cDNA Ends

Studio del DNA e dell'RNA

6.2K Visualizzazioni

article

15.12 : Metodo di sequenziamento Sanger

Studio del DNA e dell'RNA

750.4K Visualizzazioni

article

15.13 : Next-generation Sequencing

Studio del DNA e dell'RNA

85.6K Visualizzazioni

article

15.14 : Sequenziamento dell'RNA

Studio del DNA e dell'RNA

9.6K Visualizzazioni

article

15.15 : Assemblaggio e annotazione del genoma

Studio del DNA e dell'RNA

18.6K Visualizzazioni

See More

JoVE Logo

Riservatezza

Condizioni di utilizzo

Politiche

Ricerca

Didattica

CHI SIAMO

Copyright © 2025 MyJoVE Corporation. Tutti i diritti riservati