Ldlr primer

Example 2

Total RNA was isolated from mouse primary astrocytes and human primary astrocytes using RNA-Easy Qiagen kit following manufactures protocol. Semi-quantitative RT-PCR was carried out as described earlier (20) using oligo (dT) 12-18 as primer and moloney murine leukemia virus reverse transcriptase (MMLV-RT, Invitrogen) in a 20 μl reaction mixture. The resulting cDNA was appropriately amplified using Promega Master Mix and the primers for murine genes. Tfeb primer: Fwd: 5′-aacaaaggcaccatcctcaa-3′ SEQ ID NO.: 1; Rev: 5′-cagctcggccatattcacac-3′ SEQ ID NO.: 2 Ldlr primer was purchased from SantaCruz Biotechnology (Cat. No. sc-35803-PR). Amplified products were electrophoresed on 2% agarose gels and visualized by ethidium bromide staining. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) mRNA was used as a loading control to ascertain that an equivalent amount of cDNA was synthesized from each sample.

Example 2

Total RNA was isolated from mouse primary astrocytes and human primary astrocytes using RNA-Easy Qiagen kit following manufactures protocol. Semi-quantitative RT-PCR was carried out as described earlier (20) using oligo (dT) 12-18 as primer and moloney murine leukemia virus reverse transcriptase (MMLV-RT, Invitrogen) in a 20 μl reaction mixture. The resulting cDNA was appropriately amplified using Promega Master Mix and the primers for murine genes. Tfeb primer: Fwd: 5′-aacaaaggcaccatcctcaa-3′ SEQ ID NO.: 1; Rev: 5′-cagctcggccatattcacac-3′ SEQ ID NO.: 2 Ldlr primer was purchased from SantaCruz Biotechnology (Cat. No. sc-35803-PR). Amplified products were electrophoresed on 2% agarose gels and visualized by ethidium bromide staining. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) mRNA was used as a loading control to ascertain that an equivalent amount of cDNA was synthesized from each sample.