...cloning), adaptor ligation for next generation sequencing (NGS) library construction, and circularization...These applications detect specific nucleotide sequences with high specificity and quantitative accuracy...Detection Reaction (LDR), probes complementary to the sequence of interest are annealed to target DNA (genomic...fidelity thermostable DNA ligase. If the target sequence is present, the probes will ligate. Subsequent... contacts, and thus do not read specific base sequences, but are sensitive to distortions in helix shape...Gelfand. "Cloning, overexpression and nucleotide sequence of a thermostable DNA ligase-encoding gene." Gene...1756968. 7. Lauer, G., et al. "Cloning, nucleotide sequence, and engineered expression of Thermus thermophilus...

...injected animal.  PAM sequence considerations For changes 5’ to the PAM, sequences corresponding to the...developmental processes, CRISPR/Cas9 allows us to delete sequences and introduce mutations and epitopes with unprecedented... but did result in 1-2 kilobases of additional sequence being introduced. Cre-mediated excision of the...the selective cassette minimizes the sequence added, leaving a 34 bp “scar”, but currently requires additional...editing efficiency, while for edits 3’ to the PAM sequences corresponding to the opposite strand were better...they were able to insert 10 kb of non-homologous sequence or a series of single-nucleotide substitutions...an alternative gene editing system for AT-rich sequences https://doi.org/10.17912/W2237D Farboud B, Severson...

...the need for genome sequencing or annotation (5). In the event that genome sequencing can be done, this ...problem: Host binding proteins are hard to find Sequencing and cloning phage DNA can be difficult, in part...(4). To further complicate matters, even when sequenced, phage genomes tend to be choc-full of open reading...technique can also help pinpoint the genes within a sequenced genome that encode for functional cell-binding... a restriction enzyme with a short recognition sequence (like MluCI) that will cleave DNA frequently enough...Now you can grow up your colonies of interest, sequence the inserted DNA and characterize your new putative...

...insertable sequences or specific mutations, along with homologous DNA to the target sequence to be modified...and deletions along the way. HDR uses homologous sequence as a template for repair and is thus the more ...used for repair is identical to the original DNA sequence at the DSB, or it can introduce specific mutations...non-crossover events. This means all newly synthesized sequences are retained by the original damaged DNA molecule...is introduced disrupting one or both of these sequences. To prevent re-cutting and the resulting mutations...

... also directly paste sequence data into the appropriate full and partial sequence cells. *Note: If you...strain DH5α. For plasmids with highly repetitive sequences (prone to recombination in bacteria) such as lentiviral...username. Shipping address and phone number. Plasmid sequences or GenBank files. We encourage submission of QUEEN-generated...GenBank files (Mori and Yachie, 2021) but can accept sequence files in any format. Distribution status for your... amino acid change as well as any functional consequences of that change. For example, “Mutated aspartate...

... the iBARs can be detected by next generation sequencing.  In order to avoid a high false-positive rate...gRNAs and to embed these barcodes within the gRNA sequences. This allowed tracing of the performance of each...experiment by counting both the gRNAs and their iBAR sequences. Since the sgRNAiBAR library covers every annotated...stain” DNA. TALE proteins can bind specific DNA sequences which brings the attached fluorescent protein ...

...end-point sequencing assays.  This tool was designed by putting together the coding sequence of a mCherry...base edited cell populations without end-point sequencing assays. These plasmids use “Cas9-mediated adenosine...protein followed by a stop codon and the coding sequence for a green fluorescent protein (GFP). The success...

...recombineering is unsuccessful and the wild-type sequence persists, Cas9 will cleave the gene of interest...have inactivating mutations in the Cas9 or spacer sequence, and these rare events are easily detectable using...effective targeting either the promoter or the coding sequence and the template or non-template strand, among...population based on the presence of specific target sequences (Citorik et al., 2014). These could be genes, ...introduce cytosine-to-thymine substitutions in target sequences (Banno et al., 2018). These substitutions were...approximately a five-base pair window of the target sequence which could be adjusted by changing the sgRNA ...2014) Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials. Nat Biotechnol 32:1146...

...nucleases to transcription factors – to specific sequences of DNA has become commonplace. This ability has...networks. A comparable system for targeting specific sequences of RNA is highly desirable for extending the complexity...possible to produce a protein that binds an alternate sequence. By comparison, Abil explained, the specificity...

...cassettes: plant promoters, N-terminal tags, coding sequences of the gene of interest, C-terminal tags, plant...endonucleases except that they cut at arbitrary sequences a defined distance away from the recognition site...and recognition are independent. “They see one sequence and cut another,” Lohmann said. “It allows us ...