I have isolated DNA from FFPE tissue of HCC.
The DNA conc. i get is 29.6ng/ul using nanodrop
How much DNA should be required to run NGS ?
Can anyone pls help or let me know any other alternative method
The amount of DNA required for NGS (Next-Generation Sequencing) depends on the specific protocol and platform being used. Generally, NGS libraries require a certain amount of input DNA to generate enough sequencing reads for accurate analysis. For example, some Illumina protocols may require 1-2 μg of input DNA for library preparation, while other platforms or protocols may require less or more.
In your case, with a DNA concentration of 29.6 ng/μl, you can calculate the total amount of DNA you have in your sample by multiplying the concentration by the volume of your sample. For example, if you have 50 μl of sample, your total amount of DNA would be:
29.6 ng/μl x 50 μl = 1480 ng
Once you know the total amount of DNA you have, you can check the specific requirements of the NGS protocol you plan to use to determine whether you have enough DNA for library preparation. If you do not have enough DNA for your desired protocol, there are several methods for DNA amplification or enrichment that can be used to increase the amount of DNA available for sequencing, such as PCR or whole genome amplification (WGA).
It's important to note that DNA extracted from FFPE tissue may be degraded and fragmented, which can affect the quality and quantity of DNA available for NGS. Therefore, it may be necessary to perform additional quality control steps, such as fragment size analysis or library quantification, to ensure that your DNA is suitable for NGS analysis.
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