Sequence characterization of goat TLR genes

48 RESULTS Sequence characterization of goat TLR genes 4.1. Isolation and quantification of RNA Total RNA was extracted from different tissues of goat and used for cDNA synthesis to amplify goat TLR4 and TLR8 genes for cloning and sequencing. The RNA extracted by TRIzol method was further purified using RNeasy MinElute Clean-up kit (Qiagen). The concentration and purity of RNA was measured by Nanodrop (ND1000). The concentration and optical density (OD) ratio at 260 and 280nm of RNA isolated from different tissues was estimated, the values for which have been given in Table 4.1. The RNA concentration in different tissues was found to be between 564.4 ng/μl (heart) to 4984 ng/μl (intestine). For all the samples except for intestine the 260/280 ratio was around 1.9 to 2.0, indicating good quality RNA without any RNA contamination and suitable for further use. Table. 4.1. Concentration and Purity of RNA isolated from goat tissues. Tissue Conc.(ng/μl) 260/280nm Liver 4983 1.9 Lungs 4284 1.9 Kidney 3844 2.0 Skin 1082 2.0 Ovary 3513 2.0 Placenta 4876 1.9 Spleen 4975 1.87 Lymph node 4446 1.9 Heart 564.4 2.0 Mammary gland 1292 2.0 Intestine 4984 1.8 Skeletal muscle 2881 2.0