Next-generation DNA sequencing is currently limited by an inability to accurately count the number of input DNA molecules. Molecular counting is particularly needed when accurate quantification is required for diagnostic purposes, such as in single gene non-invasive prenatal testing (sgNIPT) and liquid biopsy. We developed Quantitative Counting Template (QCT) molecular counting to reconstruct the number of input DNA molecules using sequencing data. We then used QCT molecular counting to develop sgNIPTs of sickle cell disease, cystic fibrosis, spinal muscular atrophy, alpha-thalassemia, and beta-thalassemia. The analytical sensitivity and specificity of sgNIPT was >98% and >99%, respectively. Validation of sgNIPTs was further performed with maternal blood samples collected during pregnancy, and sgNIPTs were 100% concordant with newborn follow-up.

Next-generation DNA sequencing is currently limited by an inability to count the number of input DNA molecules. Molecular counting is particularly needed when accurate quantification is required for diagnostic purposes, such as in single-gene non-invasive prenatal testing (sgNIPT) and liquid biopsy. We developed Quantitative Counting Template (QCT) molecular counting for reconstructing the number of input DNA molecules using sequencing data. We then used QCT molecular counting to develop sgNIPT of sickle cell disease, cystic fibrosis, spinal muscular atrophy, alpha-thalassemia, and beta-thalassemia. Incorporating molecular count information into a statistical model of disease likelihood led to analytical sensitivity and specificity of >98% and >99%, respectively. Validation of sgNIPT was further performed with maternal blood samples collected during pregnancy, and sgNIPT was 100% concordant with newborn follow-up. 15/20

Use of green compounds to achieve the sustainable agriculture is the present necessity. Biosurfactants are one of the green compounds reported to be produced by bacteria, yeasts, and fungi which are considered to be less toxic and eco-friendly with extensive applications in pharmaceutical, cosmetics, and food industries. Bacillus group is one of the examples of biosurfactant producers. In the present study, soil isolates of Bacillus species were screened for the production of biosurfactant by oil spread assay. The activity was found positive in three isolates which were assayed for antifungal activity against the phytopathogens R. oryzae-sativae and F. solani. The biocontrol activity was observed in both extracellular and the homogenized biomass fractions of B. subtilis BP-9, B. subtilis BP-13 and Bacillus isolate PRIS-1. Different methods as acid precipitation, organic solvent extraction and salting out by ammonium sulfate were used for isolation of biosurfactant lipoeptides from the extracellular fractions of B. subtilis BP-9, B. subtilis BP-13 and Bacillus species PRIS-1. The antifungal as well as biosurfactant property was confirmed in the acid precipitated fraction of Bacillus isolate PRIS-1. The compound was identified as a high molecular weight protein by SDS-PAGE with approximate size of 100kD.

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18The genome of the budding yeast (Saccharomyces cerevisiae) has selectively retained introns in 19 ribosomal protein coding genes. The function of these introns has remained elusive in spite of 20 experimental evidence that they are required for the fitness of yeast. Here, we computationally predict 21 novel small RNAs that arise from the intronic regions of ribosomal protein (RP) coding genes in 22Saccharomyces cerevisiae. Further, we experimentally validated the presence of seven intronic small 23 RNAs (isRNAs). Computational predictions suggest that these isRNAs potentially bind to the ribosomal 24 DNA (rDNA) locus or the corresponding rRNAs. Several isRNA candidates can also interact with 25 transcripts of transcription factors and small nucleolar RNAs (snoRNAs) involved in the regulation of 26 rRNA expression. We propose that the isRNAs derived from intronic regions of ribosomal protein coding 27 genes may regulate the biogenesis of the ribosome through a feed-forward loop, ensuring the coordinated 28 regulation of the RNA and protein components of the ribosomal machinery. Ribosome biogenesis and 29 activity are fine-tuned to the conditions in the cell by integrating nutritional signals, stress response and 30 growth to ensure optimal fitness. The enigmatic introns of ribosomal proteins may prove to be a novel 31 and vital link in this regulatory balancing act. 32Keywords: small RNA, rDNA, intron, ribosome, non-coding RNA, rRNA 33 the genome, despite the loss of introns in other gene classes [4,6].