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Sorted cells were subjected to fluorescent labeling to check for purity by flow cytometry.
To check for purity the oligo was resolved on a 15% polyacrylamide gel prior to use.
The flow through was collected and analyzed in a 10% SDS polyacrylamide gel to check for purity The sample was prepared by making a dilution of protein in S200 buffer into phosphate buffer (20 mM Na2PO4, 150 mM NaCl, 12.5%, Glycerol, 1 mM DTT pH 7.4) to give a 5 µM final concentration.
After their arrival, the isolates were first sub-cultured on to sabouraud dextrose agar (SDA, BBL, Becton Dickinson Cockeysville, MD, USA) to check for purity and identifications.
Addition of the test organisms was done in all the wells except for wells of column 11 which contained neat DMSO and broth, this served as control to check for purity.
Sub-populations of cells co-expressing CD44 and CD24 at higher levels or CD24 weak/absent were collected, and a small amount of cells were re-analysed to check for purity.
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The isolates were checked for purity and preserved at −70°C for further studies.
The obtained PCR products were checked for purity and size on 1.5% agarose gel.
Cultures were checked for purity by Gram staining and observed under a microscope at 1000 ��.
Isolates obtained were further streaked on Luria Bertani (LB) agar plates and checked for purity prior to subsequent molecular analysis.
The strains were checked for purity and bacterial counts showed that 24 h incubation at room temperature in UHT milk resulted in approximately 108 cfu mL−1.
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