...different synthetic peptides corresponding to linear sequences within the middle of the human c-Myc protein or...chicken homologs. By comparing the homologous sequences in these species, they found that the N-terminal... as a recombinant antibody. Researchers first sequenced the variable domains of Myc1-9E10 in the late ...affinity for the epitope, a phenomenon known as sequence context sensitivity (Schüchner, et al. 2020). ...Myc-tag antibodies appeared to be less sensitive to sequence context.    So, do Schüchner, et al.’s results...variable epitope recognition dependent on neighboring sequence context. Sci Signal 13. https://doi.org/10.1126...

...orthologs, Cpf1 efficiency varies widely with gRNA sequence. Both groups used multiple methods to examine ...single and double mismatches throughout the 23-base sequence. Double mismatches ablated Cpf1 activity, except... they were present in the 3’ end of the target sequence (bases 19-23). Cpf1 is also sensitive to single...23. Accordingly, the 3’ end of the gRNA target sequence does not have an essential function in Cpf1-mediated...4-6 base deletions at the 3’ end of the target sequence. Cpf1’s strength may lie in its low off-target...post-HDR, repair templates should remove the PAM sequence. Many research and clinical applications require...

...discrepancies between what we sequenced in house and the submitted sequences were a concern to the plasmid... currently working on the QC/QA team as a DNA Sequencing Intern. The application process When I first...approachable.     Fig. 1: Examples of DNA sequencing QC work    In addition to my responsibilities...

...majority of information on each cancer clinical sequencing report includes rare variants that haven’t been...corresponding to cancer variants found in genome sequencing studies (as well as reference cDNA clones mapping...scientific community through Addgene. These include 634 sequence confirmed mutant cDNA clones corresponding to ... worked with many collaborators to aggregate a sequence-confirmed series of clones corresponding to known...when rare variants are discovered in clinical sequencing studies. At the Broad, my colleague Cory Johannessen...

...such as this "CRISPR 101" post. Next-generation sequencing technologies are the gold standard but they remain...assays can be designed to detect either a donor sequence integrated through homology-directed repair (HDR...designed to perfectly complement the wild type sequence and is not capable of binding a mutated target... screening tool, it still requires target DNA sequencing to confirm the exact nature of your mutations...great description of the plethora of available sequencing options can be found here. Good luck with your...

...knock-in long sequences with reduced risks of unwanted mutations in mammalian cells. Long sequences traditionally...M-CREATE) dCas9-SSAP with reduced error rate for long sequence knock-ins Visual barcodes for live cell clonal...  dCas9-SSAP with reduced error rate for long sequence knock-ins by: Lucie Wilson Le Cong’s lab has developed...SSAP will make the editing of kilobase-scaled sequences more efficient and less risky.   Fig. 3...

...clonal dynamics over time using next generation sequencing (NGS). The library was developed when Carrie ...population of cells with specifc, easily read sequences that can be used to measure their clonal abundance...barcoded clone can then be monitored over time by sequencing the barcodes in the population (all barcodes ...primers). The abundance of a particular barcode sequence then corresponds to the abundance of a specific...clone; lineage tracing is performed via barcode sequencing. Using the ClonTracer library, Bhang et al. found...

... a rapid and highly efficient way to move DNA sequences into multiple vector systems. With the appropriate...scientists can easily clone in their desired DNA sequences. Gateway technology relies on the two reactions...add attB sites to either end of the KRAS coding sequence. If you choose this strategy, it’s important to...protein expression elements (ribosome recognition sequences, start codon, stop codons, reading frame considerations...verify the integrity of your expression clone via sequencing or restriction digest! Then, you can transform...Once you generate the entry clone with your DNA sequence of interest, you can move this DNA fragment across...

... cells. Cas13 enzymes also don’t require a PAM sequence at the target locus, making them more flexible...given single base protospacer flanking site (PFS) sequence, but orthologs like LwaCas13a do not require a... target RNA from bases 12-26 of a 30 nt target sequence. For the editing portion of the protein, they ...designated this system REPAIRv1. Using next-generation sequencing, they confirmed A->I editing and found that 50...target effects observed in whole-transcriptome RNA-sequencing, the authors performed rational mutagenesis and...RESCUE further expands the RNA editing toolkit and sequences that can be edited.   As discussed above, the ...

...mariner), as a method of tagmentation for next-gen sequencing pipelines (e.g. using Tn5), and as knock-in tools...A CRISPR RNA (crRNA) comprising a 32 nt spacer sequence specifying the target site. Four proteins (TniQ...interest flanked by ~50-150 bp transposon end sequences (R and L ends).  Figure 1: (A) Components.... Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade; although 5’-CC-3’...complexes survey the genome and bind target DNA sequences using RNA-DNA base pairing. Separately, TnsABC...overnight Screen colonies phenotypically or by PCR/sequencing While CRISPR-transposons are powerful and simple...