NR-19464?? Francisella tularensis subsp. tularensis, Strain SCHU S4, Gateway? Clone Set, Recombinant in Escherichia coli, Plate 7(Clones)|Francisella tularensis subsp. tularensis|Francisella tularensis subsp. tularensis, Strain SCHU S4, Gateway? Clone Set, Recombinant in Escherichia coli, Plate 7|-80°C or colder|Pathogen Functional Genomics Resource Center at the J. Craig Venter InstituteAcknowledgment for publications should read "The following reagent was obtained through BEI Resources, NIAID, NIH: Francisella tularensis subsp. tularensis, Strain SCHU S4, Gateway
? Clone Set, Recombinant in Escherichia coli, Plate 7, NR-19464."|
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 Francisella
tularensis (F. tularensis) subsp. tularensis,
strain SCHU S4, Gateway? clone set consists of 19 plates which
contain 1693 sequence validated clones from F. tularensis
subsp. tularensis, strain SCHU S4
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%. Detailed information
about each clone is shown in Table 1.
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.
Plate viability was
confirmed for NR-19464.
Each inoculated well of the 96-well plate
contains approximately 60 ?L of E. coli
culture (strain DH10B-T1) in Luria Bertani (LB) broth containing 50 ?g/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.