Journal: BMC Neuroscience
Article Title: Glutamic acid decarboxylase 1 alternative splicing isoforms: characterization, expression and quantification in the mouse brain
Figure Lengend Snippet: Expression of GAD1 mRNA splicing isoforms and GAD2 in different mouse tissues and during development. The expression level of Isoforms 1/2 (A) , GAD2 (B) , Isoforms 3/4 (C) , Isoforms 5/6 (D) , Isoforms 7/8 (E) , Isoforms 9/10 (F) are compared by PCR amplification using mouse Multiple Tissue cDNA Panel I and cDNA from pancreas, small intestine, and large intestine as cDNA template. (G) Control PCR reaction to verify the specificity of the primers for Isoforms 1/2, 3/4 and 5/6 . In the control reaction each primer pair was tested with a plasmid containing each full length insert as a template. The template in lanes 1, 4 and 7 was plasmid containing Isoforms 1/2 as template; lanes 2, 5 and 8, plasmid containing Isoforms 5/6 ; and lanes 3, 6, 9 plasmid containing Isoforms 3/4 . Lanes (Ht) heart; (Br) brain; (Sp) spleen; (L) lung; (Li) liver; (Ms) muscle; (K) kidney; (Ts) testis; (E7) 7-day embryo; (E11) 11-day embryo; (E15) 15-day embryo; (E17) 17-day embryo; (P) pancreas; (SI) small intestine; (LI) large intestine; (N) no template control; (−) plasmid containing Isoforms 1/2 is used as template for the amplification; and Ma, marker ϕX174/HaeIII.
Article Snippet: The PCR mix (25 μl) contained 2.5 μl template cDNA, 1 μM forward and reverse primers, 200 μM dNTPs (Takara), and 0.5 U Ex Taq HS (Takara).
Techniques: Expressing, Polymerase Chain Reaction, Amplification, Plasmid Preparation, Mass Spectrometry, Marker