Surprisingly, CFC assay of the elutant
or flow-through sample containing nonadherent cells from sP tubes resulted in very few CFCs, suggesting that most of the HSPCs are captured during perfusion [average BFU-E 1 (0) and CFU-GM 12 (2)].
Shown are AC voltammograms from an E-DNA sensor before and after incubation with the saline elutant
from paper containing either the masking DNA plus 20 ng (3.
In step 1, PCR of DNA on the disks or from elutant
was amplified with primers rPLU1 (5'-TCA AAG ATT AAG CCA TGC AAG TGA-3') and rPLU5 (5'-CCT GTT GTT GCC TTA AAC TCC-3') in a standard 50-[micro]L PCR with a PTC-200 Thermocycler (MJ Research; Waltham, MA, USA) under the conditions of 94[degrees]C for 4 min, and 35 cycles at 94[degrees]C (30 see), 55[degrees]C (1 min), 72[degrees]C (1 min), with an additional extension at 72[degrees]C (4 min).
This methanol elutant
was concentrated and passed through a silica gel column using an elutropic scheme of increasing polarity.
The column elutant
was mixed with colorimetric reagent [0.
The purified RNA is electrophoresed through the gel and eluted from the bottom, the elutant
being constantly monitored by a spectrophotometer.
The capsules were submerged in PBS and 1 mL of elutant
was collected, centrifuged, and analyzed via spectrometry for 14 days.