Shotgun sequencing of UF8407
DNA was extracted from a toe pad of this specimen using a modified protocol from Soares et al. 2016 and in a lab space reserved for degraded samples. Briefly, the toe pad was rinsed in EDTA (pH 8.0) then placed in an extraction buffer containing ATL (Qiagen), proteinase K, and 1 M Dithiothreitol (DTT). The sample was incubated at 60 deg for 24 hours and then the resultant lysed sample was cleaned and concentrated with a Qiagen Nucleotide Removal Kit. DNA yield was quantified with a Qubit 2.0 Fluorometer. A DNA library was produced for UF8407 using a xGen Methyl-Seq DNA Library Prep Kit following the standard protocol but excluding the bisulfite conversion step. PCR cycles for the amplification step were 16. The library concentration was quantified with a Qubit 2.0 Fluorometer. Samples were cleaned, pooled, and shotgun sequenced on 40% of an Illumina NovaSeq, S4, using a 2x150 platform.