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BEI Resources currently limits the total number of "Made to Order" items which can be ordered by a registrant to 10 "Made to Order" items per 6 month period. Please check the Availability Status on the items you are ordering and limit orders appropriately.
NR-50781  Mycobacterium tuberculosis, Erdman Barcode Library
(Libraries)
Price: All BEI Resources products are provided
at no cost to registered researchers.
Description: Mycobacterium tuberculosis
Organism: Erdman Barcode Library
Biosafety Level: 3
Availability Status: In Stock
Store at: -80°C or colder
Contributor: S Fortune, D Schnappinger
Comments:
Quantity limit per order for this item is 5.  This item can be ordered twice a year.  Orders over this limit will be sent to NIAID for approval before shipment.

NR-50781 is a digitally barcoded Mycobacterium tuberculosis (M. tuberculosis), strain Erdman library where each bacterium in the library carries a unique, randomized sequence tag that can be used to quantitatively track the fate of each infecting bacterium.  The “barcode” consists of a random 7-mer and adjacent 75-mer library identifier tag inserted at the bacterial attB L5 insertion site of a chromosomally integrating plasmid with pJEB402 backbone. The plasmid was a standard integrating mycobacterial vector containing an Escherichia coli origin of replication, the attP element for insertion into the L5 site and is maintained by a kanamycin resistance cassette. The strain remains susceptible to all first line antimycobacterial agents once the barcode is stably inserted into the bacterial chromosome. To quantitate barcodes via sequencing, the barcode is amplified from isolated genomic DNA in two rounds of nested PCR steps.

The virulence characteristics of the barcoded M. tuberculosis have been assessed by quantitative plating and assessments of pathology in over two dozen nonhuman primates where the barcoded strain has similar virulence characteristics to the parental strain of Erdman. The complete genome of M. tuberculosis, strain Erdman was deposited in the DNA Database of Japan under accession no. AP012340.

Each vial contains approximately 0.7 mL (1.45 × 108 cfu) of M. tuberculosis, strain Erdman barcode library in Middlebrook 7H9 medium with additives (10% oleic acid-dextrose-catalase; 0.2% glycerol; 0.05% Tween® 80 and 20 µg/mL kanamycin) and cryopreservative (10% glycerol, 0.85% NaCl and 0.05% tyloxapol). The stock can be used directly for infection or can be recovered in broth culture prior to infection, depending on the lab’s infection protocol.

Note: Do not repeatedly culture the same aliquot. This can skew qtag/barcode (qbid) distribution.  Expansion of the culture is the same for all but the protocol for infection will vary depending on the route of infection. Users should follow their lab protocol once the culture is expanded. 

For-profit entities wishing to obtain this material must obtain permission from the Contributor’s institution. Please contact the Harvard University Licensing Office to obtain permission via email: hms_materialtransfer@harvard.edu.
Citations: Acknowledgment for publications should read “The following reagent was obtained through BEI Resources, NIAID, NIH: Mycobacterium tuberculosis, Strain Erdman Barcode Library, NR-50781.”
PERMITS
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  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.

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