Whole genome sequencing of Arabidopsis thaliana mutant msh1
Using picogreen concentration of msh1 sample (35ng/uL), a 20nM dilution was made as by 4.01 uL DNA and 20.99 uL TE. This was made single-stranded by 1) 5ul 20nM dilution + 5uL 0.2N NaOH, 2) wait 5 minutes, 3) add 990 uL Illumina HT1 Hyb Buffer. This created a 100pM ssDNA stock. To load the flowcell, the following were mixed: 1) 100uL 100pM stock, 2) 397 uL HT1 buffer, and 3) 3uL PhiX 10nM ssDNA control (spike-in). This yielded 500uL of a 20pM solution which was then loaded onto the MiSeq(500) flowcell.
University of Nebraska-Lincoln
msh1 rep1