title: Illumina HiSeq 2500 sequencing; Effect of Tcf7l2 Knockout in the development and identity of Mouse brain Thalamus at embryonic stage E18.5 and P60
design: Effect of Tcf7l2 Knockout in the development and identity of Mouse brain Thalamus at embryonic stage E18.5 and P60
library_construction_protocol: Whole thalamus was isolated from the brain of E18.5 mice (embryonic) either from Tcf7l2 +/+-WT or Tcf7l2 -/- KO. The mother was incapacitated by cervical dislocation. The abdomen was sliced open to locate the embryonic sac. Embryonic pups were isolated and sedated on ice immediately. Following this the pups were decapitated and the embryonic brain was sliced to reveal the thalamus. The tail of  the pup was used for genotyping the pup as Tcf7l2 wild-type (Tcf7l2 +/+) or knockout (Tcf7l2-/-). For P60, wild-type (Tcf7l2+/+)  and knockout (Tcf7l2 -/-), the mice were sacrificed, by cervical dislocation. This was followed with decapitation and isolation of brain.The for-and the hind-brain were sliced to expose the thalamus and mid-brain-cortex along with the hypothalamus and hippocampus. The cortex, hippocampus were carefully detached from the thalamus and the hypothalamus was cut out to isolate a clean sample tissue, comprising of the thalamic entity. The isolated thalamus was homogenized in Qiazol. RNA was extracted using RNeasy Lipid Tissue Mini Kit from Qiagen. The concentration of the RNA was assesed usinmg nano-drop and the quality of the RNA were assesed by visualizing the integrity of the 18s and 28s rRNA on an agarose gel. The library was prepared using TruSeq standard kit from Illumina. Before fragmentation and adapter attachment the ribosomal  RNAs were digested using RiboZero kit.
attributes.tag: Experimental Factor: genotype
attributes.value: Tcf7l2 knockout
attributes.tag: Experimental Factor: developmental stage
attributes.value: embryonic day 18