The methyl-directed DNA repair efficiency of a set of M13mp18 heteroduplexes containing 1-8 or 22 unpaired bases was determined by using an in vitro DNA mismatch repair assay. The unpaired bases of each heteroduplex residing at overlapping recognition sites of two restriction endonucleases allow independent assay of repair on either DNA strand. Our results showed that the repair of small nucleotide heterologies in Escherichia coli extracts was very similar to base-base mismatch repair, being strand-specific and highly biased to the unmethylated strand. The in vitro activity was also dependent on products of mutH, mutL, mutS, and uvrD loci and was equally efficient on nucleotide insertions and deletions. The repair levels of small heterologies were affected by base composition of the heterologies. However, the extent of repair of heteroduplexes containing small heterologous sequences was found to decrease with an increase in the number of unpaired bases. Heteroduplexes containing an extra nucleotide of 22 bases provoked very low level of methyl-directed repair.
Surface acoustic wave (SAW) sensors based on reduced graphene oxide/poly (diketopyrrolopyrrolethiophene-thieno [3,2-b]thiophene-thiophene) (rGO/DPP2T-TT) composite sensing films for the detection of ammonia were investigated at room temperature in this study. The rGO/DPP2T-TT composite films were deposited onto ST-X quartz SAW resonators by a drop-casting method. FESEM, EDS, and XRD characterizations showed that the rGO/DPP2T-TT composite film was successfully synthesized and exhibited numerous wrinkles and a rough structure, which are crucial for gas adsorption. The frequency response to 500–1400 ppb ammonia shown by the prepared SAW sensor coated with rGO/DPP2T-TT composite film increased linearly as the ammonia concentration increased. The sensor based on a rGO/DPP2T-TT composite film exhibited a positive frequency shift of 55 Hz/ppm, and its frequency response to 500 ppb ammonia was 35 Hz. The sensors thus show promising potential in detecting sub-ppm concentration levels of NH3 at room temperature, which opens up possibilities for applications in the noninvasive detection of NH3 in the breath. As a result, the rGO/DPP2T-TT composite sensor can be a good candidate for in situ medical diagnosis and indoor/outdoor environment monitoring.
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