Sentences with phrase «tagging target sequences»
The first step involves tagging target sequences with fluorescent markers.
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The GFP - tagged isoform a (plasmid pCK28 -LCB- PW01A8.1:: W01A8.1 (a) synth:: gfp:: unc - 54 3 ′ UTR -RCB--RRB- was constructed by synthesizing the W01A8.1 a sequence with modified codons to allow protection from CRISPR / Cas9 targeted sgRNA and prepared as a GeneArt ® Strings ™ DNA Fragment from Invitrogen (Invitrogen, Carlsbad, California, USA) and cloned using GeneArt ® Seamless Cloning System (Invitrogen) into pPD95.75 (NeoR).
This allows amplicons generated with targeted primers (even those tagged with the same index) to be pooled together to generate the library to be sequenced on a single flow cell.
A final ligation reaction correctly assembles the final targeting vector (Fig. 1, Step 4), and co-injection of the targeting vector and a Cas9 expression plasmid will insert the desired genetic tag and marker sequence into the targeted loci.
Like MailChimp, you can send out automatic email sequences, but CK also lets you create unique tags for each subscriber to target them for different types of marketing (different genres or email courses).