RNA isolation was performed as suggested in the Tuveson Laboratory Murine and Human Organoid Protocols, briefly the culture media was aspirated away, and the Matrigel domes were dissolved into TRIzol Reagent (Thermo Fisher), followed by the RNA isolation protocol recommended by the manufacturer. Librar...
Isolation of mouse LSK cells LSK cells were obtained using a previously described protocol12. Adult mice were euthanized, and bone marrow was extracted and passed through a 70 µm filter. Cells were centrifuged at 300g for 10 min at 4 °C, resuspended in EasySep buffer (STEMCELL, ...
The GeneJET spin columns are compatible with vacuum manifolds of standard size (e.g. Qiagen, Sigma-Aldrich, Promega). However, not all GeneJET kits are supported with a vacuum protocol. Please refer to the particular kit protocol for the detailed procedure. ...
3D). This sigma factor has previously been recognised as differentially expressed in virulent versus avirulent strains of M. tuberculosis50. Other up-regulated RNA polymerase sigma factors are sigB, sigE, sigH, sigM. Differential expression is seen among the whiB family of transcription factors, ...
The samples were costained with DAPI during the first hybridization round and the samples were imaged under Pyranose Oxidase enzyme (Sigma P4234)based anti-bleaching buffer. After imaging, the DNA probes and the hairpin polymers were digested with 1 h DNase I treatment (0.5 U / μl...
RPA is a master regulator of DNA metabolism and RPA availability acts as a rate-limiting factor. While numerous studies focused on the post-translational regulations of RPA for its functions, little is known regarding how RPA availability is controlled.
25 sections of 20 micron were collected for DNA isolation. DNA was isolated with an automated workflow (QiaSymphony) using the DSP DNA Midi kit for blood and QiaSymphony DSP DNA Mini kit for tumor samples according to the manufacturer’s protocol (Qiagen). DNA concentration was measured by Qu...
Heparin Sigma-Aldrich Cat#H3149 Neurobasal™ Medium Thermo Fisher Scientific Cat#21103049 B-27 Supplement (50X), minus vitamin A Thermo Fisher Scientific Cat#12587010 B-27 Supplement (50X), serum free Thermo Fisher Scientific Cat#17504044 Insulin solution (human) Sigma-Aldrich Cat#I9278 2-Merca...
DNA was extracted according to a general protocol. Library preparation and DNA capture were performed according to the manufacturer’s instructions. Whole-genome sequencing Genomic DNA was randomly broken into fragments to facilitate the construction of insert libraries. For human genome re-sequencing, ...
These data demonstrate the poor performance of the QuickExtract protocol when used without subsequent DNA clean-up (Ampure beads). Full size image High pure PCR template prep kit (Roche Diagnostics, Indianapolis, IN) Two hundred μL of PBS was added to the bacterial pellets and re-suspended ...