板 25(克隆），NR-1_Vaccinia virus

Note:  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.

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 Mycobacterium tuberculosis (M. tuberculosis) Gateway^? clone set consists of 42 plates which contain 3724 sequence validated clones (3294 M. tuberculosis, strain H37Rv clones supplemented with 430 unique open reading frames (ORF) from M. tuberculosis, strain CDC1551) cloned in Escherichia coli (E. coli) DH10B-T1 cells. Each ORF was recombined in vector pDONR?221 with an ATG start codon and no stop codon. The sequence was validated by full length sequencing of each entry 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.

Each 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.

Note: 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.

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 ?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.