...glycosylase inhibitor (UGI) components, which inhibit BER during cytosine-to-uracil editing, and instead introduced...

...resulted from excision by uracil N-glycosylase (UNG) during base excision repair. Adding a second copy of the...

...neuronal loop which is activated and strengthened during memory formation and memory retrieval. In neuronal...

...simultaneously measuring the activities of CaMKII and RhoA during induction of structural plasticity in single dendritic...

...bacteria, which can uptake plasmids and amplify them during cell division. Because bacteria are easy to grow...Specifically, A pairs with T and C pairs with G. During replication, DNA unwinds and the DNA polymerase...utilizes modified fluorescently tagged nucleotides. During illumina NGS, a long piece of DNA is first fragmented...Addgene all incoming plasmids are sequenced with NGS during our quality control process . NGS allows us to ...

...differ profoundly from viral particles produced during a lytic cycle. rAAV episomes can develop chromatin-like...host cell. In contrast, viral particles produced during a lytic cycle are quickly released through cell...

...are both blunt-ended, can then be fused together during a ligation reaction with T4 DNA ligase. Restriction...easy to add restriction sites to any piece of DNA during PCR amplification, allowing for it to then be digested...

...modulating the cell cycle can help as HDR takes place during the S and G2 phases. Chemically or genetically ...cytidine to uridine, which is read as thymidine during subsequent DNA repair, creating a C to T change...CRISPR Journal . 2 (6), 376–394. PMID: 31742433 Durrant, M. G., Fanton, A., Tycko, J., Hinks, M., Chandrasekaran...

...functions. Example schematic of an optogenetics procedure. A channelrhodopsin, fused to mCherry, is expressed...outside of the brain, rendering the experimental procedure less invasive. Different activation wavelengths...

...infection of 293/911 cells throughout virus production. During experiments, GFP can be used to sort cells infected...

...because they are only able to enter the nucleus during mitotic breakdown of the nuclear envelope. Lentiviruses...

...sites are called off-targets and should be examined during gRNA design. In general, off-target sites are not...

...produce full length virus from the transfer vector during the viral production stage. Many of the lentiviral...

...Carpenter JC, Snowball A, Knauss S, von Schimmelmann M, During MJ, Lignani G, Schorge S, Young D, Kullmann DM,...

...valency of 2 IgG1; IgG2; IgG3; IgG4 IgM Produced during the primary immune response; found mostly in blood...

... for all steps. Procedure Day 1 Add 200 ng DNA to each 50 µL aliquot of thawed Endura Duos on ice. Flick...representation of individual plasmids in the pooled library during amplification. This protocol is designed to be ... µL electrocompetent cells (Default: 4 tubes of Endura Duos, Lucigen, 60242-1) Alternatives include Stbl4... cm cuvette, 1.8 kV, 1 pulse. Aliquot 25 µL DNA-Endura into pre-chilled cuvette. Pulse. Caution Electroporation...contain a total of 5 mL (3 mL SOC + 2 mL transformed Endura from two separate transformations). Pro-Tip Do ...mix when removing SOC containing transformed DNA-Endura from cuvette. Shake four 14 mL Vented Falcon Tubes...pulling motion Take care not to split or gouge agar during the scraping process. Add each scrape into a 50...

...polyethylenimine (PEI) transfection protocol. This procedure can be modified for alternative packaging cell...that are below passage 15 for viral production. Procedure Seed 293T packaging cells at 3.8×10 6 cells per...DNA tube. Incubate the mixture 12–15 min at RT. During the incubation, add 10 mL of DMEM Complete to a... pellet any packaging cells that were collected during harvesting. Filter supernatant through a 0.45 μm...

...glucose, lower FBS media causes less filter clogging during harvesting. The addition of sorbitol has been shown.... Autoclave or sterile filter. Pro-Tip Stirring during the cooling period is recommended or the solution...overgrow your cells. Pass the cells twice a week during the maintenance phase and do not allow cells to.... Thaw a new vial of cells after 30 passages. Procedure Trypsinize and resuspend the HEK293T cells from...

...isolate and purify DNA fragments based on size. The procedure starts with standard agarose gel electrophoresis...with the razor blade. This is especially important during the DNA purification step, as many kits cannot ...used to determine how much of each buffer to add during the DNA isolation step. Finally, you will want ...

...content, including how-to screen captures, lab procedure protocols, and career advice videos....Video protocol guides for standard laboratory procedures. How-To Videos Screencapture walk-throughs to...Gel electrophoresis is the standard laboratory procedure for separating DNA by size for visualization and...