ClonTracer Barcoding Library
(Pooled Libraries #67267, #69830)
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Purpose
A high-complexity barcode library that “enables the high-resolution tracking of more than 1 million cancer cells under drug treatment”.
This library is offered in two sizes, depending on the number of cell lines you want to barcode.
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Vector Backbone
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Depositing Labs
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |||
|---|---|---|---|---|---|---|---|
| Pooled Library | 67267 | 2.5 µg pooled library; for amplification or to barcode 1 cell line | 1 | $540 | Add to Cart | ||
| Pooled Library | 69830 | 7.5 µg pooled library; can barcode approximately 3 cell lines | 1 | $1300 | Add to Cart | ||
† A Cas9 plasmid is NOT included with this item and will have to be ordered separately. Can be used in conjunction with pPB-R1R2_EF1aVP64dCas9VP64_T2A_MS2p65HSF1-IRESbsdpA (Addgene #113341) or any other plasmid(s) or cell lines expressing both dCas9-VP64 and MS2-p65-HSF1.
Library Details
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Library TypeBarcoding
Library Shipping
This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.
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Concentration100 ng/µL
Resource Information
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Protocols
- ClonTracer PCR Protocol (XLSX, 25 KB) NOTE! The primers in this PCR procotol have been updated since the paper was published. Please use the primers described in this protocol.
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Depositor Data
- ClonTracer v1.2 - Algorithms for deconvoluting NGS data (ZIP, 2.3 MB) NOTE! This algorithm software contains minor bug fixes and a new “library_mode” geared toward analyzing plasmid library complexity.
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Articles Citing this Pooled Library
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Terms and Licenses
Academic/Nonprofit TermsIndustry Terms- Not Available to Industry
Trademarks- Zeocin® is an InvivoGen trademark.
Depositor Comments
Current stock of the library can be used for experiments of less than or equal to 1 million total cell infections.
NOTE: Lentiviral vectors can recombine during library amplification, resulting in an ~1 kb band containing the origin and ampicillin resistance cassette. This recombination product should not adversely affect library function as it does not contain lentiviral packaging sequences or the PuroR gene and will not be selected during experiments.
A recombinant band has been detected in the parent stock of ClonTracer. The concentration of DNA on our website reflects the concentration of the unrecombined plasmid pool and can be used for barcoding as described in the associated protocol.
The proportion of the recombinant band in the pool will increase upon subsequent amplifications. Gel purification will eliminate most of this recombinant band (see example gel) and does not alter barcode representation. If the sample is to be used for amplification, we recommend starting with a larger prep (gigaprep) from the original sample or gel purifying the amplified library to remove the recombinant band and performing another amplification with the purified sample.
Figure 1: Diagram of ClonTracer barcoding process.
These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
ClonTracer Barcoding Library was a gift from Frank Stegmeier (Addgene #67267)
ClonTracer Barcoding Library was a gift from Frank Stegmeier (Addgene #69830) -
For your References section:
Studying clonal dynamics in response to cancer therapy using high-complexity barcoding. Bhang HE, Ruddy DA, Krishnamurthy Radhakrishna V, Caushi JX, Zhao R, Hims MM, Singh AP, Kao I, Rakiec D, Shaw P, Balak M, Raza A, Ackley E, Keen N, Schlabach MR, Palmer M, Leary RJ, Chiang DY, Sellers WR, Michor F, Cooke VG, Korn JM, Stegmeier F. Nat Med. 2015 May;21(5):440-8. doi: 10.1038/nm.3841. PubMed