The CloneTracker™ 5-Barcode and 10-Barcode Cell Labeling Kits (Cat.#s BCRPP5-V and BCRPP10-V) provide packaged lentiviral particles for labeling 5 and 10 different cell populations/clones, respectively, with unique barcodes which differ in sequence but are all the same size. The barcode constructs in these kits also contain a red fluorescent protein marker (TagRFP) and puromycin-resistance selection marker.
The CloneTracker™ 4-Barcode-Plus Cell Labeling Kits (Cat.# BC4P-V, BC4PPO-V) both provide 4 different packaged barcode constructs, each of which differ in both size and sequence and can be differentially amplified by PCR. The barcode constructs in Cat.# BC4P-V express both TagRFP fluorescent protein and puromycin-resistance selection marker, but those in Cat.# BC4PPO-V express only puromycin-resistance selection marker.
This Manual provides instructions on how to titer and transduce a target cell population with a single lentiviral barcode construct with the purpose of generating a cell population uniformly labeled with a single genomically integrated unique barcode sequence. The differently barcoded cell populations (e.g. with different genotypes) can be mixed together and used for in vitro or in vivo experiments, for example to monitor proliferation rate of different clones with and without treatment conditions (e.g. drug). The relative proliferation rate of individual barcode-labeled groups of cells can be identified in a mixed population by standard sequencing or, in the case of the CloneTracker Barcode-Plus Cell Labeling Kit, by gel electrophoresis analysis of the amplified barcodes.
The CloneTracker Cell Labeling Kits are designed for labeling only a limited number of different cell populations. For studying clonal dynamics and growth heterogeneity in the complex cell population with mixed representation of different cell types (e.g. different by phenotype or genotype), we recommend Cellecta’s CloneTracker™ 50M Lentiviral Barcode Library. The CloneTracker lentiviral library comprises more than 50 × 106 different barcodes and can be effectively used to measure representation of different clones (e.g. stem cells, drug-resistant clones) by Next-Generation Sequencing (NGS) on the Illumina platform. Furthermore, the cells labeled using the BCRPP5 or BCRPP10 kits can be mixed together with cells labeled with the CloneTracker 50M Barcode Library as they have different unique barcode sequences but can be amplified and sequenced by the same set of primers.
The experimental design for using barcoded cell populations is mainly defined by your model system and depends on your specific scientific goals. Puromycin-selected barcoded cell populations can be mixed together and analyzed for relative growth characteristics under different conditions in both in vitro and in vivo. For example, the mixed barcoded cell populations (e.g. with different genotype) can be mixed with other cell types (e.g. stromal cells) and/or treated with different agents (e.g. drugs) or grown under different growth conditions (e.g. hypoxia vs normoxia, attached vs suspension, etc). Alternatively, the single barcoded cell population(s) can be spiked-in to heterogeneous cell populations transduced with the CloneTracker 50M Barcode Library, and these single barcoded cells could be used as an internal positive/negative control to monitor the growth characteristics of other cell types present in the heterogeneous cell population.
Please read the entire user manual before proceeding with your experiment. Also, please note that, when working with pseudoviral particles, you should follow the recommended guidelines for working with Biosafety Level 2 (BSL-2) materials.
The purchase of all Cellecta Products is covered by Cellecta’s standard Terms and Conditions of Sale as described on Cellecta’s website, and selected Products containing particular technology or having certain features are also subject to restrictions on use as outlined in the Label License section of our website. Please review these Terms and Label License Restrictions before opening and using your Product and, if you are not able to abide by the restriction, contact Cellecta to return the item to Cellecta for a full refund.
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