Biological lithography: Improvements in DNA synthesis methods
Locations of the errors by diffraction. (a) Aerial image calculated by Optolith . (b) Hybridization of mixed base 25-mer. fluorescence microscope. (c) Fluorescence signal of monomer (dT). The entire region is deprotected and directly coupled with Cy3 phosphoramidite. (d) Cross section of (a). The arrows indicate the location of diffraction peaks. (e) Cross section of (b). (f) Cross section of (c). Notice the revelation of hidden strand sites by hybridization.
DNA synthesis Monte Carlo simulation of mixed-base 25-mer oligonucleotides and its experiment. (a) Monte Carlo simulation: 2D distribution of probe oligomers at the center pixel of array. The line represents the edges of a DMD micromirror. (b) Central cross section of (a). (c) Hybridization signal of mixed-base 25-mer. fluorescence microscope. (d) Central cross section of (c).
(a) Radiachromic film exposure “without image locking:” The outer square reference frame was exposed at , after which each pixel of 4 lines was consecutively exposed every 10 min. At 21st and 33rd pixels, environmental temperature was set to change as marked. (b) Same radiachromic film exposure as (a) “with image locking” in operation. (c) Hybridization of 25-mer synthesized “without image locking.” fluorescence microscope. (d) Same synthesis as (c) “with image locking.”
Hybridization of mixed-base 90-mer. Probe sequence (3’-ATCACGCAAG GACATGTATT GGAAGCCCGT ACCGTGAGAA CTTTTTCAGA ACGGCAAAGA ATACTCTGGT CCCACCAAGT ACTACTACTG-5’).
Inverted capping. (a) Fluorescence signal of monomer (dT). Only patterned region (60 half-life times exposure) is coupled with Cy3 “without inverted capping.” (b) Fluorescence signal of monomer (dT). The same synthesis as (a) but “with inverted capping.” (c) fluorescent microscope image of (b). (d) Aerial image calculation for inverted mask by Optolith.
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