BI 117 Recitation Session 1. Techniques in Developmental Biology. Part 1 : Methods Using DNA, RNA, and Protein Jon or jev Kerckhoff 017 [email protected] The life codes. Methods with DNA and RNA. DNA->RNA->Protein - the flow of biological information. DNA = A, T, C, G. D: deoxyribose.
Coding sequence of SpBra
Non-coding sequences (intergenic and intronic) in the vicinity of SpBra
Regulatory element binding sites
Purpose: to generate a superabundance of copies of your DNA fragment
Cloning is useful for many downstream applications!
Substrate for Alkaline phosphatase
PHow it Works
Often conjugated to a flourescent molecule
Embryos were stained at room temperature with the following primary antibodies: mouse monoclonal anti-Ftz, mouse monoclonal anti-Engrailed (a gift of N. H. Patel), and rabbit polyclonal anti-Even-skipped (a gift of M. Frasch). The primaries were visualized with Cy3 anti-mouse and FITC anti-rabbit
Genetics. 2004 September; 168(1): 161–180.
Immunoprecipitation (IP)The technique of precipitating a protein antigen out of solution using an antibody that specifically binds to that particular protein.Purpose: to isolate a specific protein from a lysate or crude extract
Histone H1 accumulation in sktl salivary glands. Salivary glands from wild-type (wt) and sktldrosophila larvae were dissected, analyzed by SDS-PAGE, and immunoblotted with histone H1 and tubulin antibodies. (Top) Antihistone H1 immunoblot. Histone H1 antibody recognizes both the phosphorylated (32 kD) and nonphosphorylated (31 kD) forms of histone H1. Nonphosphorylated histone H1 is completely absent in sktl mutant salivary glands. (Bottom) Same blot probed with tubulin antibody as a loading control.
From: Genetics, Vol. 167, 1213-1223, July 2004
Kuroda et al., 2005
Nature Reviews Genetics