title: NextSeq 500 paired end sequencing; Targeting NAT10 enhances healthspan and lifespan in a mouse model of human accelerated aging syndrome.
design: Targeting NAT10 enhances healthspan and lifespan in a mouse model of human accelerated aging syndrome.
library_construction_protocol: Mice were euthanized by CO2 inhalation followed by cervical dislocation.  Upon dissection the mouse tissues were snap-frozen in liquid N2 and immediately stored at -80ºC. RNA was extracted from tissues from n>5 independent mice/group using the RNeasy fibrous tissue mini kit (50) (cat. No.7404; Qiagen) and quantified using the NanoDrop 1000 Spectrophotometer (Thermo Fisher Scientific). 2μg RNA/sample was used to produce cDNA using the High-Capacity RNA-to-cDNA kit (cat. No. 4387406; Applied Biosystems/Thermo Fisher Scientific). qPCR was carried out using the TaqMan system (Universal Master Mix II, with UNG, 4440038; Applied Biosystems/Thermo Fisher Scientific) on an Applied Biosystems Quant Studio 3 machine. n≥3 mice were used for each genotype, with 50ng cDNA for each sample run in triplicate or quadruplicate. Thermo Fisher Scientific Nat10 (Mm00462302_m1) and Cdkn1a/p21 (Mm00432448_m1) primers were used as experimental probes while Gapdh (Mm99999915_g1), Actb (Mm00607939_s1) and/or Rn18s (Mm03928990_g1) were used as endogenous controls. The Relative Standard Curve pre-set programme was used throughout and data analysis was performed using the Relative Quantification application, powered by the Thermo Fisher Scientific cloud platform. Illumina: TruSeq Stranded mRNA Library Prep Kit (Cat. No. RS-122-2101)
attributes.tag: Experimental Factor: age
attributes.value: 10
attributes.tag: Experimental Factor: compound
attributes.value: none
attributes.tag: Experimental Factor: disease
attributes.value: Hutchinson-Gilford progeria syndrome
attributes.tag: Experimental Factor: phenotype
attributes.value: NAT10 knockout