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Unfortunately, this fosmid was chimeric, which likely occurred as a by-product formed during genomic library production from the ligation of two noncontiguous fragments.
Barcoding the product before library construction provides a system of multiplexing with minimal library production costs.
A protocol was developed to tag PCR products with 5' 6-base oligonucleotide barcode extensions before pooling for sequencing library production using standard Illumina adapters.
For the library production and the Sav expression, the E. coli strains TOP10 and BL21 (DE3) are used, respectively.
For library production, 107 HEK-293T cells were seeded per 15 cm dish the day before transfection.
Investments in compound management infrastructure and chemistry library production capability allowed significant growth in the size, diversity and quality of the BMS compound collection.
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A fosmid genomic library was constructed from approximately 1.1 µg of DNA from the 0.2 5 µm plankton fraction using the CopyControl™ Fosmid Library Production Kit (Epicentre) as described by the manufacturer's instructions.
The fosmid library was created using the CopyControl Fosmid Library Production Kit with the pCC1FOS vector (Epicentre).
Additionally, a fosmid library was constructed with the Copy Control Fosmid Library Production Kit (Biozym Scientific, Hess. Oldendorf, Germany).
Selected fosmid clones were grown in lysogeny broth medium + chloramphenicol and multicopy fosmid production induced as described by the manufacturer of the CopyControl Fosmid Library Production Kit (Epicentre).
8μg of B2O viral DNA was used in large-insert fosmid library construction using the Epicentre CopyControl Fosmid Library Production Kit (CCFOS110) as previously described [ 44].
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