What are the gDNA requirements for the Axiom 2.0 assay?
All human Axiom arrays (except the Axiom Genome-Wide Pan-African Array Set) require a total of 100 ng.
The Axiom Genome-Wide Pan-African Array Set requires a total of 300 ng or 100 ng per array.
Diploid plants and animals require 150 ng per array and polyploid plants and animals require 200 ng per array.
For Axiom Microbiome Arrays, a total of 50 ng of gDNA or 17.5 µL of cDNA reaction + 2.5 µL reduced TE buffer starting material is required per array.
Please refer to the Axiom 2.0 gDNA Sample Preparation Quick Reference for more details. Additionally, you can refer to pages 12-13 of the Axiom 2.0 Assay 96-Array Format Manual Workflow User Guide.
Quality recommendations:
- Starting gDNA must be double-stranded to support accurate concentration determination.
- gDNA must be of high purity.
- DNA should be free of DNA polymerase inhibitors. Examples of inhibitors include high concentrations of heme (from blood) and high concentrations of chelating agents (i.e., EDTA).
- The gDNA extraction/purification method should render DNA that is generally salt-free because high concentrations of particular salts can also inhibit enzyme reactions.
- DNA purity is indicated by OD260/OD280 and OD260/OD230 ratios. The OD260/OD280 ratio should be between 1.8 and 2.0 and the OD260/OD230 ratio should be greater than 1.5.
- DNA must not be degraded.
The approximate average size of gDNA may be assessed on a 1% agarose gel using an appropriate size standard control.
Approximately 90% of the DNA must be greater than 10 Kb in size. Control DNA can be run on the same gel for side-by-side comparison.
We recommend that DNA samples that do not meet these criteria be cleaned up as described under Genomic DNA Cleanup in the Axiom 2.0 Assay 96-Array Format Manual Workflow User Guide, pages 15-17.
Find additional tips, troubleshooting help, and resources within our Microarray Analysis Support Center.
The Axiom Genome-Wide Pan-African Array Set requires a total of 300 ng or 100 ng per array.
Diploid plants and animals require 150 ng per array and polyploid plants and animals require 200 ng per array.
For Axiom Microbiome Arrays, a total of 50 ng of gDNA or 17.5 µL of cDNA reaction + 2.5 µL reduced TE buffer starting material is required per array.
Please refer to the Axiom 2.0 gDNA Sample Preparation Quick Reference for more details. Additionally, you can refer to pages 12-13 of the Axiom 2.0 Assay 96-Array Format Manual Workflow User Guide.
Quality recommendations:
- Starting gDNA must be double-stranded to support accurate concentration determination.
- gDNA must be of high purity.
- DNA should be free of DNA polymerase inhibitors. Examples of inhibitors include high concentrations of heme (from blood) and high concentrations of chelating agents (i.e., EDTA).
- The gDNA extraction/purification method should render DNA that is generally salt-free because high concentrations of particular salts can also inhibit enzyme reactions.
- DNA purity is indicated by OD260/OD280 and OD260/OD230 ratios. The OD260/OD280 ratio should be between 1.8 and 2.0 and the OD260/OD230 ratio should be greater than 1.5.
- DNA must not be degraded.
The approximate average size of gDNA may be assessed on a 1% agarose gel using an appropriate size standard control.
Approximately 90% of the DNA must be greater than 10 Kb in size. Control DNA can be run on the same gel for side-by-side comparison.
We recommend that DNA samples that do not meet these criteria be cleaned up as described under Genomic DNA Cleanup in the Axiom 2.0 Assay 96-Array Format Manual Workflow User Guide, pages 15-17.
Find additional tips, troubleshooting help, and resources within our Microarray Analysis Support Center.