...genes replaced.  Additionally, 2A peptides could be PCR-cloned between your genes of interest and you can...

... individual gene edited clones are identified by PCR. Then, if necessary for subsequent experiments, the...

...Overall, it worked. We pulled it off! Check out the PCR Primer video! Learn: Producing videos like this would...

...targeting region from the vector. Alternatively, PCR-generated or commercially synthesized linear dsDNA...

... What is a Plasmid? How to Design Primers for PCR Agarose Gel Electrophoresis Inoculating Liquid Bacterial...

...analyzers, and any ultra sensitive equipment (RT-PCR, Microscopy, Fluorescence Scopes, Flowcytometers,...

...restriction enzyme (RE) recognition site, RE analysis of PCR product could be used to detect success. Although...

...generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...biology 11.1 (2015): 31-37. pCri system The Gomis-Ruth lab designed the pCri system to simplify the protein...the repository: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit FREQ Seq...depending upon your specific protein folding needs. The pCri system has recently been used to study a matrix ....8 (2015): 4728-4740. You can find more info on pCri in Mary Gearing’s blog post on the system Easing...Easing the Protein Purification Process with pCri Cell-free protein expression kit Cell-free expression ...

...Addgene Blog Verify Your Constructs with Colony PCR Read about Tags for Protein Purification Resources... use by the Rinehart lab: pCRT7 NT TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a ...68292 C321.ΔA bacterial strain – Addgene #68306 pCRT7 NT TOPO E17TAG GFP plasmid - Addgene #68295 SupD...

...expressed them along with Cas9 in the T-cell line 2D10. PCR amplification and Sanger sequencing of pooled samples...

..., SacI restriction site and a linker region. The PCR product is then ligated into the MCS/SacI site in...can occur in this system if a small contaminating PCR product formed from primer dimers and containing ...clone. ORFs from an ORFeome Gateway library or any PCR-amplified ORF cloned into pDONR221 can be expressed...

... repair templates using Addgene plasmids. Design PCR primers to add T7 promoter-left homology arm and ...

... showed correctly sized inserts following colony PCR of 10-20 colonies using primers bordering the multiple...

...and the sequence of gene of interest in separate PCR amplication reactions for recombination in SLIC cloning...the ccdB cassette, which is not copied during the PCR amplification step, so that only vectors with the...or multiply phosphorylated proteins (B40 OTS and pCRT7-GFP). To demonstrate the improvements of the system...

...Plasmids 101: Origin of ReplicationPlasmids 101: PCR...

...such as physically carrying out experiments like PCR and running gels, but in research you also experience...

...vectors using BsaI, and promoter-gRNA fragments are PCR amplified for cloning into one of three Zeamays codon-optimized...

...and these rare events are easily detectable using PCR. The system also functions in S. pneumoniae and can...created two “pCRISPomyces” systems for use in Streptomyces (Cobb et al., 2015). pCRISPomyces-1 includes ...array, while pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays...pCas9: carries Cas9 and chloramphenicol resistance pCRISPR: carries a spacer targeting the gene of interest...machinery are first electroporated with pCas9. Then, pCRISPR is introduced along with an oligonucleotide repair...advantages including shorter plasmid curing time. pCRISPomyces for editing in Streptomyces  Streptomyces bacteria...

...annealed synthetic oligos - eliminating the need for PCR or additional molecular cloning steps during plasmid...occurred at higher frequency and could be detected by PCR while large deletions were relatively rare. The vectors...target G-protein coupled receptors (GPCRs) yet one third of the GPCRs in the human genome are orphan receptors...Mammalian Expression 74334 pCR8-Magneto2.0-p2A-mCherry Gateway 74333 pCR8-Magneto2.0 Gateway 74307...are unknown. Identifying new ligands for orphan GPCRs is of significant medical interest, however efforts...allows researchers to interrogate the druggable human GPCR-ome using a quick, affordable, and ubiquitous reporter...recruitment assay incorporating 314 codon-optimized GPCR sequences for human cell line expression. In a twist...

...controlling GPCR activation have been developed to selectively study individual types of GPCRs with greater...be used for lower frequency stimulation. GPCR signaling GPCR signaling is a complicated process involving...change the membrane potential. OptoXRs are modified GPCRs. When exposed to light, they activate downstream... coupled receptors  G-protein coupled receptors (GPCR) regulate cellular function through multiple intra-cellular...have been developed for utilizing light to control GPCR activation and downstream signaling cascades (reviewed...Levitz, 2020), including light-sensitive chimeric GPCRS (OptoXRs) (Tichy et al., 2019), photoswitchable ...rhodopsin, and the intracellular domain of an endogenous GPCR, such as adrenergic receptors. When exposed to light...