To gain some insight into absorption into voids of complex geometry, and to provide a representation of heterogeneities in surface interaction energy, the cylindrical throats were converted to double cones.
Macrophages were harvested for RNA which was converted to double stranded cDNA, labeled with Cy3 and 4 µg of labeled cDNA hybridized to Musmusculus 4×72 whole genome array.
Briefly, first RNA was converted to double stranded cDNA followed by a clean-up step using spin columns.
An excess of cycles was used during this step to ensure that all of the single-stranded oligo was converted to double stranded template.
Briefly, 500 ng of total RNA was converted to double stranded cDNA using T7-oligo dT) primers followed by IVT7-oligo dTto primers biotinylatedcRNA.
Briefly, a total of 5 μg RNA was converted to double - stranded cDNA using the one-cycle cDNA Synthesis Kit (Affymetrix, Inc., Santa Clara, CA) and T7-Oligo (dT) Primer.
Briefly, total RNA from three L. sigmodontis samples (adult male, adult female, microfilaria) was converted to double stranded cDNA using Evrogen's MINT cDNA synthesis kit.
Briefly, mRNA was converted to double stranded cDNA in a PCR reaction containing random hexamer primers, Superscript III Reverse Transcriptase (Invitrogen) and supplied buffer.
Per sample, 2 μg of total RNA spiked with bacterial RNA transcript positive controls (Affymetrix, Santa Clara, CA, USA) was converted to double stranded cDNA.
Poly A+ RNA was isolated using the poly A Spin™ mRNA isolation kit (New England Biolabs, Beverly, MA). 5 μg of polyA+ RNA was converted to double stranded cDNA using the cDNA Synthesis Kit (Life Technologies, Gaithersburg, MD).
