... a ~20 nucleotide targeting sequence and ~75 nucleotide scaffold sequence) is consistent between plants...contain a plant pol III promoter and gRNA scaffold sequence and allow researchers to insert targeting oligos...is commonly tagged with a nuclear localization sequence to enhance targeting to the nucleus, and several...the specifics - promoters used, precise protein sequences or domains, and methods of delivery - the underlying...

...repetitive tRNA sequences must be avoided, different tRNAs with optimized 5’ leader sequences can be used ...plant source), by tuning the length of 5’ leader sequences of a given tRNA to achieve the highest efficiencies...

...protocol before performing single-cell RNA sequencing. The sequencing results covered TFs representing 92% ...average of 206 cells per TF ORF. From this large sequencing endeavor, a map of possible TF-induced cellular...

... (bEBPs) at a sequence upstream of the promoter called the upstream activating sequence (UAS). The Wang...phenotype can be identified by next-generation sequencing of the genomic DNA. However, some cell types ...

...transcriptional activator that binds to UAS enhancer sequences found in DNA. It then recruits transcription machinery... genes and siRNA encoded downstream of the UAS sequence are only expressed when Gal4 is expressed. Gal4...multiple genes under the control of UAS enhancer sequences. For example, if you discovered five new genes...

.... Piecing together multiple PCR products using sequence homology is also common. Overlap extension PCR... to splice together two PCR amplicons based on sequence homology, and the subsequent product is then digested...fragment ends have secondary structure or too much sequence similarity. REPLACR-mutagenesis: Letting bacteria...

...inducible lambda-red gene, and an IPTG-inducible sgRNA sequence targeting the pMB1 replicon on pTargetF (Jiang... this system, you’d first introduce your sgRNA sequence into the pTargetF construct and then introduce... process begins with the addition of the sgRNA sequence in pEcgRNA. Next both plasmids are transformed...pTargetF, we replaced the original 20 bp specific sequences on pTargetF with the ccdB gene containing BsaI...

... strand at each replication fork. Termination sequences present in the chromosomal DNA ensure that the...pSC101 itself. RepA binds to directly repeated sequences called iterons located near the ori and directly...ColIb-P9 plasmid, is complementary to part of the sequence forming this pseudoknot, so its binding prevents...replication may differ in the specialized proteins or sequences necessary for initiation or the mechanisms that...

...platform is compatible with enrichment, single-cell sequencing, and optical pooled screens in human cells. Addgene... improve the specificity of ssAAV, a targeting sequence of miR124T was inserted into the vector to silence...alone or in combination with miR124T targeting sequences, provides a simple yet valuable tool for microglia... REVeRT to functionally reconstituting coding sequences for a range of applications, including CRISPR ...

...longer stretches of sequence could be used. However, a practical limitation is sequencing depth and cost. ...variants with optimal function. High-throughput DNA sequencing is then used to measure the frequency of each...more codons you test in one library, the more sequencing reads you will need. See Fowler et al for more...

...: shRNAs are short RNA sequences which target complementary mRNA sequence for degradation. Viral introduction...viral vector for your gene of interest – an shRNA sequence specific to your gene’s mRNA or a sgRNA targeting...

...in your organism or cell type? You can use DNA sequencing or other molecular cloning techniques to determine...resistance gene followed by the target site (the same sequence you’d like to mutate in the genome) and an out-of-frame.... A gRNA that targets your desired gene (whose sequence is also found in the reporter gene). A control...

...reconstituted in vivo to form a protein identical in sequence to the unmodified enzyme. Each half of the base... cell. Intein splitting removes any exogenous sequences and regenerates a native peptide bond at the split...markers, transformation acceptor vectors, and sequences for genomic integration by homology directed repair...

... a protospacer flanking sequence, so you can target virtually any RNA sequence! In bacteria, Cas13d-mediated...The Cas13d proteins they identified have little sequence similarity to previously identified Cas13a-c orthologs...

...working on lab operations software for next-gen sequencing (NGS), and high-throughput screening. While some...in the US to learn where the gaps are in their sequencing workflows, any problems they’re having with our...region. She is particularly interested in DNA sequencing tech, and the intersection of technology, software...

...These vectors contain an Autonomously Replicating Sequence (ARS) derived from the yeast chromosome. As the...part of an ARS along with part of a centromere sequence (CEN). These vectors replicate as though they ...DNA (apparently independent of a known specific sequence) dictate the replication of these vectors. S. ...

...HHCS-associated sequences had lower IREB2 binding enrichment compared to wild-type sequences and that loss...however, knock-in of BoxB sites into endogenous DNA sequences could allow one to study RNAs expressed from their...

...lowering Cas9 expression, and truncating gRNA sequences used for targeting; however, these options can...the gRNA (see figure below) Because off-target sequences have less complementarity to the gRNA, they have...Lab similarly thought that if they could weaken sequence independent interactions between Cas9 and DNA,...

... available, and the collection of samples and sequencing is being organized and funded by someone else...the makerspace comfortable with next-generation sequencing data and the bioinformatics pipeline. It’s possible... the Illumina GAIIx we recently acquired  for sequencing again - it’s too inefficient and the reagents...

...also the first eukaryotic genome to be entirely sequenced and is very amenable to genetic manipulation. ...organisms, the C.elegans genome is also completely sequenced and over 60% of human genes have an ortholog in...Kerstin, et al. "The zebrafish reference genome sequence and its relationship to the human genome." Nature496.7446...extrachromosomal maintenance and integration of transforming sequences." The EMBO journal 10.12 (1991): 3959-3970. PubMed...