Transposase mediated construction of RNA-seq libraries.

RNA-seq has been widely adopted as a gene-expression measurement tool due to the detail, resolution, and sensitivity of transcript characterization that the technique provides. Here we present two transposon-based methods that efficiently construct high-quality RNA-seq libraries. We first describe a method that creates RNA-seq libraries for Illumina sequencing from double-stranded cDNA with only two enzymatic reactions. We generated high-quality RNA-seq libraries from as little as 10 pg of mRNA (∼1 ng of total RNA) with this approach. We also present a strand-specific RNA-seq library construction protocol that combines transposon-based library construction with uracil DNA glycosylase and endonuclease VIII to specifically degrade the second strand constructed during cDNA synthesis. The directional RNA-seq libraries maintain the same quality as the nondirectional libraries, while showing a high degree of strand specificity, such that 99.5% of reads map to the expected genomic strand. Each transposon-based library construction method performed well when compared with standard RNA-seq library construction methods with regard to complexity of the libraries, correlation between biological replicates, and the percentage of reads that align to the genome as well as exons. Our results show that high-quality RNA-seq libraries can be constructed efficiently and in an automatable fashion using transposition technology.

Cyanoacrylate vs. sutures in clean and clean-contaminated surgical wounds - a randomised control study.

Objectives: Various techniques of closure of surgical incisions have been described ranging from various suture materials, staples and tapes to adhesive compounds. Cyanoacrylate is an adhesive compound available for surgical incision closure. Although sutures have been the preferred universal choice for surgical incision closure, glue is gaining popularity in specific places like pediatric injuries, facial injuries, laparoscopic incision closure, etc. This study aimed to compare the results between the application of cyanoacrylate and conventional suturing. Methods: In this randomized control study, patients were divided into two groups of 100 each. The surgical incisions were closed using cyanoacrylate glue in Group A patients and polyamide (EthilonTM 2-0) in Group B patients. Post-operative pain was assessed using Visual Analogue Scale on the first, third, and seventh day. The wounds were evaluated for complications on post-op days 1, 3, 7, and 30 using the ASEPSIS score. Cosmetic outcome was assessed at the end of first month using the Modified Hollander Cosmesis Scale. Results: Post-operative pain was significantly less in the glue group on days 1, 3, and 7. Wound infection with dehiscence occurred in 4 cases (4 %) in Group A and one patient (1 %) in Group B, which was statistically insignificant. There was no significant difference in cosmetic outcomes in either Group. Conclusions: Cyanoacrylate is a good alternative to sutures in skin closure of clean and clean-contaminated surgical wounds.