...phosphate backbone modifications The recognition sequence for CRISPR-Cas9 targeting is led by a ~20 bp gRNA...CRISPR RNA (crRNA). In addition to the 20 bp guide sequence, the crRNA also contains a ~20 bp repeat region...validated gRNA repository to find pre-tested gRNA sequences for your next CRISPR experiment.    Chemical...which is photocaged and unable to engage a target sequence. Upon a brief light exposure (several seconds)...

...Yamagata et al used previously published nanobody sequences that targeted the following antigens: GFP, RFP...actin-binding protein gelsolin, but any nanobody sequence could be used to make a RANbody. RANbodies are...secreted from cells due to their IgGκ signal peptide sequence, making them easy to harvest from cell culture...useful RANbody, consider leaving a link to its sequence in the comments below. References Sanes, J.R....

...whole exome sequencing of embryo derived-ES cells.They also conducted whole genome sequencing in selected...rates to untreated controls. Ma et al. performed sequencing of 23 potential off-target editing sites in the...off-target effects, they limited their whole genome sequencing to a small percentage of edited embryos. Since...

...with a 1.9 kb stuffer sequence that can be removed and replaced with an shRNA sequence of your choice. More...libraries and can be combined with high-throughput sequencing to perform multiplexed analysis of cells pre-...backbones) here. Cloning and expressing new shRNA sequences pLKO (Plasmid #10878) This plasmid has been used...

...adding long-read sequencing capabilities to complement our next-generation sequencing, setting new standards... We’re also building a portal to easily share sequences and characterization data for over 150,000 plasmids...

...containing the modifying sequence flanked by approximately 50 base pairs of sequence homologous to the chromosomal... typically contain both a protein modification sequence and a selectable gene product for isolation of...

... reaction for traditional restriction cloning. Sequence and Ligation Independent Cloning (SLIC) could ...Invitrogen and based on lambda phage integration sequences, relies on site-specific recombination reactions...components and define their final order through sequence homology. Similarly, Golden Gate assembly allows...FastCloning: a highly simplified, purification-free, sequence-and ligation-independent PCR cloning method." ...

... the exact original sequence, it can restore the original protein-coding sequence using a recoding strategy...synthetic sequence will only spread in transgenic laboratory populations with that sequence2. Both methods...; a subsequent drive can restore the wild-type sequence (save for the residual sgRNAs and possibly cas9...unpredictable ecological effects. Because the consequences of such a mistake would necessarily extend far...

...immunoprecipitation followed by high-throughput sequencing, ChIP-Seq, is the go-to method for mapping where...noise, reducing the read depth necessary when sequencing and lowering the amount of tissue needed per ...subsequent library preparation and high-throughput sequencing. Improvements to the CUT&RUN method and using...

...Transposons are sequences of DNA that can move around in a genome. In a laboratory setting, transposons...the tranposon to integrate. Additionally, the TA sequence is duplicated near the gene of interest insertion...between a greater number of extinct transposon sequences and then testing various combinations of changes...