ClonTracer Barcoding Library
(Pooled Libraries #67267, #69830)

• 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.

• Vector Backbone:

  pRSI9-U6-(sh)-UbiC-TagRFP-2A-Puro

• Depositing Labs: Frank Stegmeier
• Publication: Bhang et al Nat Med. 2015 May;21(5):440-8. doi: 10.1038/nm.3841.

Library Details

• Library Type: Barcoding

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.

• Concentration: 100 ng/µL

Resource Information

• Protocols:
  • ClonTracer Library Transformation (DOCX, 18.8 KB)
  • 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.
  • Using the barcoding library (DOCX, 48.3 KB)
• 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.
• Articles Citing this Pooled Library:
  • ClonTracer 2.5 µg (#67267)
  • ClonTracer 7.5 µg (#69830)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA
• Evrogen Limited Use Label License for FPs

Industry 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.

Diagram of ClonTracer barcoding process