Project description:20 random DNA barcodes were designed in silico and transfected into PC3 cells. Barcodes were sequenced using Illumina-Miseq technology to find the sequence and their respective copy numbers. Current file contains the raw data of these DNA barcodes in fastq format
Project description:20 random DNA barcodes were designed in silico and transfected into PC3 cells. Barcodes were sequenced using Illumina-Miseq technology to find the sequence and their respective copy numbers. Current file contains the raw data of these DNA barcodes in fastq format Validating an algorithm called SRiD that generates random DNA barcodes that do not match a genome of interest, in this case human genome. 20 DNA barcodes were used for this validation.
Project description:Proof of principle and testing experiments on iPS2-sci-seq have been performed on two batches of hPSC-derived cardiomyocytes at day 23. Cells have been treated with Tetracycline throughout differentiation. For this experiment it was used a 2-level of indexing sci-RNAseq protocol (Cao J. et. al, Science 2017). During RT and PCR indexing steps, additional primers have been used to enrich for UCI and shRNA barcodes for the iPS2-seq pipeline. Sequencing has been performed on a NextSeq 500 with a High Output kit 75 cycles. Read1, 18 cycles; Index1, 10 cycles; Index2, 10 cycles, Read2, 52 cycles.
Project description:Embryonic neocortical cells were isolated from E14.5 WT mice, followed by TrypLE Express treatment and trituration to generate a single-cell suspension. Plasmid DNA was introduced into primary neocortical cells using Neon Transfection System. Next, neocortical neurospheres were cultured in serum-free media containing, then control and Prdm16-overexpressing cells were corrected after GFP sorting of 1 day cultures. We found that marked upregulation of modulators of mitochondrial metabolism and ROS balance in Prdm16 GOF cells
