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To obtain the dipolar broadening effect or inter-spin distances, spectra for the S90R1 and S117R1 singly-labeled mutants and the S90R1-S117R1 doubly-labeled mutant were acquired at physiological temperature with increasing concentrations of glycerol (Fig. 2A D).
SEM images of upper developing leaves, showing mature trichomes of WT and cvy1 mutant were acquired at different magnifications as previously described [ 30].
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Orco 1, ninaB P315, Dcp-1 RNAi, drice RNAi, Histone H2AvD-GFP, and the Matα GAL4 mutant strains were acquired from the Bloomington Drosophila Stock Center (strain numbers 23129, 24776, 28909, 32403, 35518, and 7063, respectively).
pCS2+-Myc/cyclin E harboring wild-type and mutant forms (T380A and T380S mutants) of cyclin E were acquired from Clurman Lab (Fred Hutchinson Cancer Research Center, Seattle, WA, USA).
Planktonic cultures of wild type E. coli and single gene knockout mutant ΔgadX and ΔuspE were acquired from overnight cultures in 0.1×LB.
Single amino acid mutants (T127A or S127D) were acquired by a similar strategy.
Specifically, for each spin-labeled single-cysteine mutant, a pair of 2D H- N Hspectractra were acquired at a protein concentration of 200 µM in 40 mM H-DPC: one for the spin-labeled sample in the paramagnetic form, and another after adding ascorbic acid (to 10 mM) to the sample to reduce the nitroxide, yielding the diamagnetic sample.
The microarray contained four subarrays in which four genotypes were acquired by mixing two mutant types and two wild types.
Data were acquired from wyEx5968 and wyEx6022 transgenes coexpressed in the ptrn-1 tm5597) ptrn-1 tm5597
Fly stocks were acquired from the Bloomington Stock Center, with the exception of brca2 KO (see below, Creation and isolation of mutants).
Others were acquired.
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