The kit utilizes the robust Q5 Hot Start High-Fidelity DNA Polymerase along with custom mutagenic primers to create insertions, deletions and substitutions in a wide variety of plasmids. After PCR, the amplified material is added directly to a unique Kinase-Ligase-DpnI (KLD) enzyme mix for ...
Aberrant translation causes ribosome stalling, which leads to the ubiquitination of ribosomal proteins and induces ribosome-associated quality control. As part of this quality control process, the E3 ubiquitin ligase RNF10 monoubiquitinates ribosomal pro
RBM10-EGFP was generated by amplifyingRBM10from pFRT-TO-RBM1026, followed by double digestion of the PCR Product withNheI andEcoRI (NEB) and ligation intoNheI/EcoRI linearized pEGFP-N3 vector (Clontech) using T4 DNA ligase (NEB). The tet-on lentiviral plasmids carryingRBM10-EGFP were con...
DNA was released from chromatin by adding 5 μL of cleanup buffer (900mM NaCl, 300 mM EDTA, 1.1% SDS, 4.4 mg/mL Proteinase K (NEB)) followed by an incubation for 30 min at 40°C. Tagmentated DNA was isolated using 2 × volumes of SPRI beads and eluted in 21 μl. For library...
Baselines are adjusted to zero for all curves and double-referenced by subtracting a sensorgram of buffer injected over the coated surface from the experimental sensograms to give curves representing specific binding Curves are modeled assuming a simple 1:1 interaction to generate the equilibrium and...