Author: Long-Cheng LiSource: Protocol OnlineAbstract: Simplified method for preparing G+A ladder run along with footprinting reaction. It's much simple than the original Maxima-Gilbert sequencing rea ...
DNaseI FootprintintSolutions10X Binding Buffer200 mM Tris 8.0 200 ml 1M Tris pH 8.0500 mM NaCl 100 ml 5M NaCl10 mM EDTA 20 ml 0.5 M EDTA pH 8.0680 ml Qstore at room temperatureDNaseI Dilution Buffe ...
Author: Long-Cheng LiSource: Protocol OnlineAbstract: Simplified method for preparing G+A ladder run along with footprinting reaction. It's much simple than the original Maxima-Gilbert sequencing rea ...
Principles of nucleic acid hybridization5.2.1. Nucleic acid hybridization is a method for identifying closely related nucleic acid molecules within two populations a complex target population and a co ...
Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as single-stranded or double-stranded molecules (see F ...
Nucleic acid hybridization assays using cloned target DNA and microarray hybridization technologySome of the technologies described in the preceding section (e.g. Southern blot hybridization and dot-b ...
Nucleic acid hybridization assays using cloned DNA probes to screen uncloned nucleic acid populationsNumerous applications in molecular genetics involve taking an individual DNA clone and using it as ...
Nucleic acid hybridization assays using cloned DNA probes to screen uncloned nucleic acid populationsNumerous applications in molecular genetics involve taking an individual DNA clone and using it as ...
In a microcentrifuge tube prepare a solution of linear DNA (25-50ng) in deionized water or TE buffer (10-35µl). Add: 10X ligation buffer 5µl 50% PEG 4000 solution (for blunt ends only) 5µl deionized ...
DNA labeling by nick translation reagents: DNA for labeling (concentration c 150 ng/µl) modified nucleotides: Biotin-16-dUTP Digoxigenin-11-dUTP conc. 1nmol/µl (Boehringer Mannheim) dNTPs (regular nu ...
DNA labeling by nick translation reagents: DNA for labeling (concentration c 150 ng/µl) modified nucleotides: Biotin-16-dUTP Digoxigenin-11-dUTP conc. 1nmol/µl (Boehringer Mannheim) dNTPs (regular nu ...
Linker Ligation (with T4 DNA Ligase) In a microcentrifuge tube prepare a solution of blunt ended dephosphorylated DNA (100-500ng) in TE buffer (5-7µl). Add 1-2µg of phosphorylated linkers in 5µl of TE ...
De-phosphorylation of DNAThe preferred method of de-phosphorylation uses the buffer system described by Pharmacia for most of their restriction endonucleases. You will need: 10 x OPA+ Buffer (100mM Tr ...
Thermal InactivationA simple reversible way to a stop restriction reaction is by adding EDTA which chelates Mg2+ thereby preventing catalysis. If further manipulations of the digested DNA are to be pe ...
The restriction/modification system in bacteria is a small-scale immune system for protection from infection by foreign DNA. W. Arber and S. Linn (1969) Plating efficiencies of bacteriophage lambda (l ...
Partial Endonuclease Digestion Prepare a 100 µl reaction mixture containing the DNA of interest in the appropriate 1X restriction enzyme buffer. Divide the reaction mixture between five prechilled mic ...
Restriction digestionRestriction enzyme digestions are performed by incubating double-stranded DNA molecules with an appropriate amount of restriction enzyme in its respective buffer as recommended by ...
Restriction enzymes are expensive ($40 to $200 a vial): keep the enzymes at -20 C. Plan your digests to be small and convenient. The digest is composed of DNA sample (volume should not be more than a ...
Restriction digestionRestriction enzyme digestions are performed by incubating double-stranded DNA molecules with an appropriate amount of restriction enzyme in its respective buffer as recommended by ...
Materials: Restriction enzymes of choice such as BamH1 and EcoRI Restriction enzyme reaction buffer such as MULTI-CORE (TM) (Promega) 70 % Ethanol 100 % Ethanol 3 M Sodium acetate (pH 5.2) Distilled w ...
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