DNA制备

Southern Analysis Procedure (for analysis of genomic DNA or ordering of clones): 1) Prepare genomic DNA (ref.p.9.22 Maniatis) from cultured cells using one T75 flask (wash two times with PBS and then ...

Non-radioactive Probes I. Via random hexamers 1. Solutions:10X hexa nt miundefined: 500 mM Tris-Cl pH 7.2100 mM MgCl21 mM dithioerythritol (DTE)2 mg/ml BSA62.5 A260 units/ml (1.56 mg/ml) random hexanucleotid ...

RNA/Zeta Probe Dot Blotting protocol1)Make up RNA (up to 20μg)dissolved in sterile H2 OTE or 0.5% SDS to 500μl with ice-cold sterile 10mM NaOH1mM EDTA and apply it to Zeta Probe membraneheld in ...

The standard solution typically used for both pre-hybridisation and hybridisation is based on that given in Maniatis et al.(1982)with both Denhart's solution and heterologous DNA being replaced by hep ...

About 1/2 to 1 cm of tail should be cut from properly marked mice (using toe or ear clipping etc) about the age of 2-3 weeks. Place these tails into marked 1.5 ml Eppindorf tubes for processing; they ...

1. Depurination of DNA fragments: Wash gel in 0.25 M HCl 5' (small gels) 7' (large gels)2. Denaturation: Wash gel in (0.5 M NaOH 1.5 M NaCl) for 20' (small) to 30' (large).3. Neutralization: Wash in ( ...

SolutionsProcedure Design complementary oligonucleotides with compatible half sites on the ends (I use BamHI and BglII). Dry down 300 ng of the two purified oligos together and resuspend in 9 m l Q wi ...

Protocol for Annealing OligonucleotidesOligo Name:Lot Number:Total nmol:Volume of Annealing Buffer added:Oligo Name:Lot Number:Total nmol:Volume of Annealing Buffer added:Annealing Buffer: 10mM Tris p ...