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275-3-2 (OPN expressing) cells were cultured in DMEM with 3% FBS and tested for mycoplasma: positive cells were treated with Lookout Mycoplasma Elimination Kit (Sigma, St Louis, MO, USA).
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A mutation was introduced into the DES hQC expression vector using Unique Site Elimination Mutagenesis kit (Amersham Pharmacia Biotech).
Experimental elimination of c-kit in mice mobilized with plerixafor or anti-VLA-4 resulted in a negative effect of mobilization [ 61].
Computational modeling of the experimentally identified mutations showed that these mutations reduced the binding affinity through one of the three scenarios: through SCF destabilization, through elimination of favorable SCF/c-Kit intermolecular interactions, or through allosteric changes.
Total RNA was recovered from the gDNA elimination reaction using the RNeasy Micro kit (Qiagen) RNA clean up protocol.
The kit also includes elimination of genomic DNA step prior to reverse transcription. 1 μg of total RNA was used for cDNA preparation.
It is supposed that this effect can be maintained even after elimination all of implanted c-Kit+ cells.
Zambuto said the funds will go toward three of the foundation's main initiatives: healing programs, educational work and advocacy work ― the latter of which focuses on the elimination of the U.S.'s rape kit backlog. .
Poly(A)+RNA (0.9 5.0 µg each) was used for cDNA synthesis (Just cDNA Double-Stranded cDNA Synthesis Kit, Stratagene, CA) after elimination of residual contaminating genomic DNA using the Turbo DNA-free Kit (Applied Biosystems, Ambion, CA).
Subsequent depletion of bacterial ribosomal RNA yielded an RNA sample enriched for bacterial transcripts (MICROBExpress, Applied Biosystems, Ambion, CA), which was subjected to cDNA synthesis (Just cDNA Double-Stranded cDNA Synthesis Kit, Stratagene, CA) after elimination of residual contaminating genomic DNA using the Turbo DNA-free Kit (Applied Biosystems, Ambion, CA).
This kit contains a gDNA-elimination step to avoid gDNA contamination.
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