Name | Manufacturer | Cat. # | |
Nuclei Fixation Kit v2 | Parse Biosciences | ECF2003 | |
Nuclei Extraction Buffer | Miltenyi Biotec | 130-128-024 | |
RNase Inhibitor, murine | New England Biolabs | M0314L | |
PBS | HyClone | SH30256.02 | |
Debris Removal Solution | Miltenyi Biotec | 130-109-398 | |
7.5% BSA | Life Technologies | 15260037 | |
gentleMACS C Tube | Miltenyi Biotec | 130-093-237 | |
gentleMACS Octo Dissociator | Miltenyi Biotec | 130-095-937 | |
MACS SmartStrainers (70 um) | Miltenyi Biotec | 130-110-916 | |
MACS SmartStrainers (30 um) | Miltenyi Biotec | 130-098-458 | |
pluriStrainer (20 um) | pluriSelect | 43-50020-03 | |
NucBlue Fixed Cell ReadyProbes | Thermo Fisher | R37606 | |
Millicell Disposable Hemocytometer | Millipore | MDH-2N1-50PK | |
Mr. Frosty | Sigma-Aldrich | 635639 |
Name | Reagent | Volume (for 8 samples) | Final Concentration | |
Lysis buffer | Nuclei Extraction Buffer | 35 ml | NA | |
40 U/ul RNase inhibitor | 175 ul | 0.2 U/ul | ||
PBS | PBS | 35 ml | NA | |
HBSS | HBSS | 20 ml | NA | |
Debris Removal Solution (DRS) | Debris Removal Solution (Miltenyi) | 8 ml | NA | |
NB-BSA + RNase inhibitor | Nuclei Buffer (Parse Biosciences) | 3.15 ml | NA | |
7.5% BSA | 350 ul | 0.75% | ||
RNase inhibitor (Parse Biosciences) | 44.1 ul | |||
NB + RNase inhibitor | Nuclei Buffer (Parse Biosciences) | 5 ml | NA | |
RNase inhibitor (Parse Biosciences) | 44.1 ul | |||
RSB (x 2 aliquots!) | PBS | 24.6 ml | NA | |
7.5% BSA | 333 ul | 0.1% | ||
RNase inhibitor | 125 ul | 0.2 U/ul |