We narrowed to 432 results for: sequence

...issue is sequence similarity. If fragments in a multicomponent assembly have 5′ or 3′ sequence homology...If cloning methods had personalities, SLIC (sequence- and ligation-independent cloning) would be a true...reliable cloning method, but it is limited by its sequence constraints. The long overhangs must not contain... often limited to specially designed plasmids. Sequence- and ligation-independent cloning: A SLIC makeover...depositor Stephen Elledge. His new method, named sequence- and ligation-independent cloning (SLIC), eliminated...and linearized vectors must have homologous DNA sequences (20–60 bp) on either end and single-stranded 5...multicomponent assembly, as fragments with overlapping sequence homology can be assembled in a unique order with...

... long asymmetric core spacer sequence. The asymmetry in the core sequence gives the loxP site directionality...the canonical loxP sequence is ATAACTTCGTATA-GCATACAT-TATACGAAGTTAT. The loxP sequence does not occur naturally...in P1) sites are 34-base-pair long recognition sequences consisting of two 13-bp long palindromic repeats...inserting loxP sites at deliberate locations in a DNA sequence allows for very specific manipulations as discussed...or on separate pieces of DNA. Both 13bp repeat sequences on a single loxP site are recognized and bound...: If the sites face in the same direction, the sequence between the loxP sites is excised as a circular...indicates which bases may vary from the canonical sequence. Among these are loxN (GtATACcT), lox2272 (GgATACtT...

...target sequence) in my HR template an issue? A8: Yes. If you have intact protospacer+PAM sequence within... to mutate the PAM 'NGG' sequence in the HR template. Since the PAM sequence is required for successful...from the Zhang group. Q2: Should I add the PAM sequence to the oligo when cloning my target spacers into...backbone? A2: There is no need to add the NGG PAM sequence. Because the 'NGG' of the PAM is used to select...Q3: When designing oligos for cloning my target sequence into a backbone that uses the human U6 promoter...to add a G nucleotide to the start of my target sequence? A3: The human U6 promoter prefers a 'G' at the...adds it when they order the oligo. If your spacer sequence starts with a 'G', you naturally have one and ...

...: The transposon is the DNA sequence that moves around. This DNA sequence encodes the proteins the transposon...the protein that’s required for the transposon sequence to move from one DNA site to another. This protein...Different transposons insert at different DNA sequences or target sites. Integration of most transposable...results in the duplication of this target site sequence at the site of insertion.  Types of transposons...behind the CAG DNA footprint from its terminal sequences at the excision site after cleavage by the transposase...transposons, it does not leave behind a DNA footprint sequence after excision. piggyBac can mobilize DNA over...Beauty and piggyBac, Tol2 has a weak consensus sequence for its preferred target integration site: TNA...

... site donor and acceptor sequences were tested and with the optimal sequences, the group achieved ~80%... non-endogenous sequences, such as a 77bp long highly recombinogenic homology sequence derived from the...trans-plicing, splice site donor and acceptor sequences are used to reconstitute the two pieces of the...step, but optimization of the exon-splitting site sequence can enhance the rate of trans-splicing (Lai et..., the 5’ half of the transgene, a splice donor sequence and the region of homology. The AAV encoding part...contains the region of homology, a splice acceptor sequence, the 3’ half of the transgene. The full length...mechanism. The splice site donor and acceptor sequences and the homologous region of the two vectors can...

...localized to specific loci based on the guide RNA sequence. A year later, they developed CRISPRainbow (Ma...set of gRNA scaffolds that contain two hairpin sequences (either MS2, PP7, or boxB) that each recruits ... CRISPRainbow labeling. Two MS2, PP7, or boxB sequences inserted into the sgRNA generates blue, green,... labeling. A combination of MS2, PP7, or boxB sequences in the same sgRNA will generate additional colors...living cells. Broccoli aptamers are nucleic acid sequences that bind to the small molecule, DFHBI-1T to induce...of the success of the MS2 and PP7 RNA hairpin sequences in CRISPRainbow, they knew they could be good ...created another sgRNA containing the PP7 hairpin sequence (sgRNA-Sirius-8XPP7) to recruit GFP to the loci...

... Guide Sequence Identify a TAM site (5′-TTGAT-3′) in your target gene. Choose a 20-nt sequence immediately...guide insertion by Sanger sequencing using either 92F or M13R as the sequencing primer. Step 7: Transformation...) near the target DNA, TnpBs also need a short sequence motif called a TAM (transposon-associated motif...located just downstream (3′ end) of the target sequence, while for Cas12a and TnpB, the motif is found...Deinococcus radiodurans (Dra2TnpB) recognizes the TAM sequence 5′-TTGAT-3′, while TnpB from Deinococcus geothermalis...right-end element, and a 20-nucleotide (nt) guide sequence that is complementary to the DNA target. The 20... 20-nt guide region binds to the DNA sequence immediately downstream of the TAM. This RNA-DNA hybrid formation...

...achieved by the sequencing of plasmids isolated from blue or white colonies. Sequencing of blue colonies...showed the expected complete sequence for the lacI/lacZ genes (1), while sequencing of white colonies showed...overhang sequences under standard reaction conditions. This data set allows quantitation of sequence-dependent...restriction enzymes and careful choice of junction sequences guided by experimentally-derived DNA Ligase fidelity...indicating successful reconstruction of the coding sequence for beta-galactosidase in the lacI/lacZ cassette... Gate assembly through application of ligation sequence-dependent fidelity and bias profiling." bioRxiv... Potapov, Vladimir, et al. "A single-molecule sequencing assay for the comprehensive profiling of T4 DNA...

...homologous sequence to the right and/or left of the knock-in site. The length of that sequence and the makeup...Knock-in mutations typically require an exact DNA sequence to be incorporated at a precise genomic location...introduce the edit. Finally, you must stipulate what sequence to introduce, which is done through the design...to the targeted locus on either side of the new sequence. This homology will guide the cell to use the ...which will result in the incorporation of your sequence, a.k.a a knock-in! So how does the donor get used... can efficiently knock-in a few hundred bps of sequence. However, if you have a larger knock-in (200 bp...al., Nat Chem Biol). Mutate the PAM or sgRNA sequences Cas9 likely goes through multiple rounds of cleavage...

...interest: The transgene sequence is flanked by long terminal repeat (LTR) sequences, which facilitate integration...the transfer plasmid sequences into the host genome. Typically, it is the sequences between and including...Leukemia Virus) or MSCV (Murine Stem Cell Virus) sequences. Packaging genes (viral Gag-Pol): Gag is a structural...virus post‐transcriptional regulatory element; sequence that stimulates the expression of transgenes via...the 5' and 3' ends of the provirus). Contains sequences necessary for activation of viral genomic RNA ...the addition of a poly A tract just after the R sequence. This post was written and updated by multiple...