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Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment.The methicillin-resistant Staphylococcus aureus (S. aureus), strain COL Gateway® clone set consists of 25 plates which contain 2343 sequence validated clones from S. aureus strain COL cloned in Escherichia coli (E. coli) DH10B-T1 cells. Each open reading frame was constructed in vector pDONR™221 (Invitrogen™) with a native start codon and no stop codon. The sequence was validated by full length sequencing of each clone with greater than 1X coverage and a mutation rate of less than 0.2%.Information related to the use of Gateway® Clones can be obtained from Invitrogen™. Recombination was facilitated through an attB substrate (attB-PCR product or a linearized attB expression clone) with an attP substrate (pDONR™221) to create an attL-containing entry clone. The entry clone contains recombinational cloning sites, attL1 and attL2 to facilitate gene transfer into a destination vector, M13 forward and reverse priming sites for sequencing and a kanamycin resistance gene for selection. Please refer to the Invitrogen™ Gateway® Technology Manual for additional details.Every inoculated well of the 96-well plate contains approximately 60 µL of E. coli culture (strain DH10B-T1) in Luria Bertani (LB) broth containing 50 µg/mL kanamycin supplemented with 15% glycerol.
Production in the 96-well format has increased risk of cross-contamination between adjacent wells. Individual clones should be purified (e.g. single colony isolation and purification using good microbiological practices) and sequence-verified prior to use. BEI Resources does not confirm or validate individual mutants provided by the contributor.
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biological materials provided by BEI Resources, please select this link: Compliance Requirements.
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