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"We are looking for substances that use the cell lines and methods described in the patent to find new anti-HIV drugs that work by binding to CCR5 receptors and blocking them," said Dr. Haseltine, who did pioneering work in sequencing the genome of HIV at Harvard's Dana-Farber Cancer Institute.
Our findings allow evidence-based choices of Cas9-expressing lines and methods for generating knock-in alleles.
Description of cell lines and methods for Western blot can be found in Additional file 3. The cytotoxic effect of methotrexate treatment of PBMCs and cell lines was estimated using a cytotoxicity detection kit (LDH; Roche, Mannheim, Germany).
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Extensions corresponding to these states are predicted using polymer models of elasticity (black dots and dotted lines, 'Materials and methods').
Using this value as threshold, we genotyped 438,915 markers in each of the 10 DH lines ('Materials and methods') and could confidently assign genotypes to 3,672,610 loci.
Future analyses using more cell lines and different methods of separating migratory from stationary cells may yield new findings.
The Nanos2-knockout mouse lines and PCR methods used for verification of mutant alleles have been described previously (Tsuda et al., 2003).
Oftentimes, these methods are categorized by the type of laser scanner they operate on, namely methods for line scanners and methods for 2.5-D laser scanners.
Tcm metacyclic trypomastigotes were incubated overnight with cells from each cell line (Materials and Methods).
Relaxing curves (red) are obtained after SSB dissociation, and are well fit to a polymer model of bare DNA (black dotted line, 'Materials and methods').
The generated HSC-like cell lines were denoted dox-dependent hematopoietic progenitor cell (DoxHPC) lines (see Materials and Methods and ref. [47]).
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