American Society of Hematology

Fig. 1.

$Fig. 1. Schematic for production of the normalized hematopoietic progenitor cDNA library and construction of microarray gene chips. / At top, single-strand circles represent the starting cDNA library produced by means of filamentous phage rescue. The ssDNA was hybridized with driver (10 000 IMAGE consortium cDNAs) with appropriate blocking oligonucleotides. The fraction that remains single stranded (flowthrough from HAP column) was converted to double-strand circles, electroporated into DH10Bα, and propagated under ampicillin selection to generate an amplified normalized cDNA library. Large-scale sequencing of clones was performed with the use of the M13AEK forward primer. To make the myeloid-specific gene chips, the sequenced cDNA clones were amplified by PCR, purified, and printed onto polylysine-coated glass slides.$

Schematic for production of the normalized hematopoietic progenitor cDNA library and construction of microarray gene chips.

At top, single-strand circles represent the starting cDNA library produced by means of filamentous phage rescue. The ssDNA was hybridized with driver (10 000 IMAGE consortium cDNAs) with appropriate blocking oligonucleotides. The fraction that remains single stranded (flowthrough from HAP column) was converted to double-strand circles, electroporated into DH10Bα, and propagated under ampicillin selection to generate an amplified normalized cDNA library. Large-scale sequencing of clones was performed with the use of the M13AEK forward primer. To make the myeloid-specific gene chips, the sequenced cDNA clones were amplified by PCR, purified, and printed onto polylysine-coated glass slides.

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