Chromatin lysates were subjected to column-based DNA clean-up (BioBasic, BS664, Markham, ON, Canada). Pre-IP input lysate from each cell line was used to generate a standard curve by creating 5 samples of 10-fold dilutions with known amounts of chromatin (50-0.005 ng). Two microliters of each sample was mixed with 8 µL of Fast SYBR™ Green Master Mix (Sigma-Aldrich, #4385612, St Louis, MI, USA) and the respective primer pairs, and subsequently run on an ABI ViiA7 qPCR system. Samples were run in triplicates. For downstream analyses, the chromatin amount present per sample was derived from input-generated standard curves and fold enrichment was calculated for SS18-SSX immunoprecipitation against IgG control. Primer sequences designed to target the promoter sequences are listed in Table 3 .13 (link),17 (link)
Bs664
Manufactured by Bio Basic
Sourced in Canada
The BS664 is a compact benchtop centrifuge designed for general laboratory applications. It features a maximum speed of 6,000 RPM and a maximum RCF of 4,000 x g. The centrifuge can accommodate a variety of rotor types and sample volumes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using bs664
1
Chromatin Quantification and Enrichment Analysis
2
Nucleosome Array Digestion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Two micrograms of reconstituted nucleosome arrays were incubated with 0.2 micrograms recombinant protein for 10 minutes at room temperature in 1X NEB4 buffer (New England Biolabs). AvaI restriction enzyme (12U per timepoint) was added and the reaction quenched on ice in 80% PCR cleanup binding buffer. Timepoints were subjected to PCR cleanup (Bio Basic, cat#BS664) and visualized via 0.7% agarose gel electrophoresis in 0.2X TB.
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