Use 50 ng gDNA as input. Amplicon sizes of 150 bp and 175 bp are supported.
Quanitfy and dilute the DNA to the input amount of 50 ng. You can dilute and store more than the required DNA for later use. Dilute DNA in RS1 and SS1 and store as described in the Quantify and Dilute DNA section of the TruSeq Gentoype Ne Reference Guide (document # 1000000033138).
Quantify the starting genomic material using a fluorescence-based quantification method, such as a Qubit dsDNA Assay Kit or PicoGreen. Do not use a UV-spectrometer-based method.
Fluorescence-based methods employ a dye specific to double-stranded DNA (dsDNA) and specifically and accurately quantify dsDNA, even when many common contaminants are present. In contrast, UV spectrometer methods based on 260 OD readings can overestimate DNA concentrations due to the presence of RNA and other contaminants common to gDNA preparations.