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The 3D grid of the pore space binary 1 and the 3D grid of the solid space binary 2 images were multiplied to the Swirr or Sor, and dry sample sets to isolate the fluid in the pore space area, and the mixed beads in the solid area, respectively.
To test the feasibility of sorting beads on the basis of C12-resorufin fluorescence, we mixed beads coated with anti-HA antibodies and ∼1000 molecules of CpI DNA per bead with a 50∶1 excess of beads coated with antibodies but no DNA.
Aliquots of 25 μl of mixed beads were then added to all wells.
All wells then received a further 25 μl of lysis buffer followed by 25 μl of mixed beads.
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In brief, 25 µl of pre-mixed beads coated with capture antibodies were added to a 96-well plate.
Following digestion, samples were immobilized to the SPRI beads via addition of well-mixed beads at 3× concentration, then a wash was performed as described in the Broad Institute protocol.
Figure 8 displays sections of the mixed-bead pack relative to residual fluid structures.
The average porosity for the mixed-bead pack is comparable to that of the glass bead and plastic bead packs where their average porosity was 41.6 and 42.1 %, respectively.
Moreover, the saturation history of oil might affect the amount of residual oil trapped after the water injection, as the amount of oil saturation of the acrylic bead packs (91.68 %) was higher than that of mixed-bead packs (85.92 %), resulting in higher residual oil saturation.
Plasma samples diluted 1 2,500 with dilution buffer (30 μL/well) or standards (30 μL/well) along with 10 μL of biotinylated antigens in blocking buffer were incubated with 10 μl of pre-mixed bead sets into the wells of a pre-wet 96-well microtitre plate.
Technical issues specific to 454 technology include: nucleotide calling difficulties within homopolymers; sequencing failure arising from incomplete homopolymer extension; base mis-incorporation by residual nucleotides during the nucleotide flow step; mixed template beads, and overrepresented single templates distributed across different beads [ 34].
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