Micrococcal Urinary tract infections in young women.
Drop-ChIP
Micrococcal nuclease (MNase), detergents, DNA barcodes, and sample DNA are combined using a microfluidic method to form nucleosomal regions flanked by DNA barcodes.
Using site-directed mutagenesis and the CRISPR-associated protein 9 (CRISPR/Cas9) system, they engineered DNA fragments carrying a variety of clinically important alterations, including the EGFR T790M, L858R and exon 19 deletion, KRAS G12D mutation, and more complex EML4-ALK rearrangements and mixed them with
micrococcal nuclease-digested HEK293 cell-line DNA to obtain DNA samples with varying allelic fractions of mutant DNA.
(39) By
micrococcal nuclease (MNase) assays to examine chromatin accessibility, (40) we revealed that the region around the STAT3 binding site in the Gfap promoter was digested well in lgNSCs, which are capable of Gfap expression with LIF stimulation, whereas this region in both WT and TKO ESCs cells and in mgNSCs was protected from the digestion (Fig.
Each well was then treated with 200 units of
Micrococcal Nuclease (MNase; Sigma-Aldrich) and 1 [micro]M of SYTOX Green (Gibco, Invitrogen) and incubated for 10 minutes at room temperature in the dark.
Micrococcal nuclease (Takara Bio, Shiga, Japan) was added to a final concentration of 30 U/mL and incubated for 30 min at room temperature with occasional homogenization.
RPMI (1 mL) containing
micrococcal nuclease (MNase) (1U/[micro]L) was then added to each well to digest NETs at 37[degrees]C for 20 min.