Mali P, Aach J, et al. (2013) Barcoding cells using cell-surface programmable DNA-binding domains. Nat Methods, 10(5):403–406.
Using IDT gBlocks® Gene Fragments
to generate several surface zinc-finger DNA-binding domains (sZFs), this research group created cell lines that expressed these distinct domains on their surface. The scientists could then essentially barcode the different cell lines by exposing the cells to fluorophore-conjugated dsDNA in a multistep labeling approach. (Fluorophore-conjugated DNA probes
were provided by IDT and contained 4 phosphorothioate bonds at both the 5’ and 3’ ends to enhance protection against nucleases prevalent in extracellular medium.) The live cells could be tracked, and also specifically isolated and enriched from within a heterogeneous cell mixture using dsDNA probes conjugated to magnetic beads or on dsDNA arrays (amine-conjugated oligonucleotides
used on the arrays were provided by IDT). The researchers envision a broad range of synthetic biology applications for sZF-expressing cells including multiplex tracking of gene activity to study gene networks and pathways.
gBlocks® Gene Fragments are double-stranded, sequence-verified, DNA genomic blocks, 125–2000 bp in length, that can be shipped in 2–5 working days for affordable and easy gene construction or modification. gBlocks Gene Fragments have been used in a wide range of applications including CRISPR-mediated genome editing, antibody research, codon optimization, mutagenesis, and aptamer expression. They can also be used for generating qPCR standards. With gBlocks Gene Fragments, IDT provides a tool that accelerates your research and makes synthetic biology easier and more accessible than ever, for any lab.
Learn more about gBlocks Gene Fragments at www.idtdna.com/gblocks.
Read how gBlocks Gene Fragments have been applied in other research projects:
Towards next generation biosensors—See how fluorescent protein-based Ca2+ indicators based on naturally occurring substrates are assembled using gBlocks Gene Fragments. These sensors are being developed to monitor in vivo neural activity.
Site-directed mutagenesis—improvements to established methods—Site-directed mutagenesis techniques have relied primarily on PCR and standard cloning methods. Read about some of the common cloning methods used for mutagenesis and how double-stranded DNA fragments (gBlocks Gene Fragments) can save you both time and money.
Easily-Designed Standard Curves for qPCR—Use this easy way to combine control templates/multiple targets onto a single construct, and the advantages that it can provide for PCR experiments.
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