Protocol for Dnase I footprinting. Includes: Labeling of probe; Synthesis of probe by PCR; Dnase I foot printing. - [Read Dnase I Footprinting Protocol]
Dnase-chip: A High Resolution Method to Identify DnaseI Hypersensitive Sites using Tiled Microarrays. This method can be applied globally or in a targeted fashion to any tissue from any species with a sequenced genome. - [Read Dnase-chip Protocol]
Protocol describes how isolated nuclei are incubated with varying amounts of Dnase I. Genomic DNA is then isolated from the nuclei and digested with a restriction enzyme, analyzed by gel electrophoresis, and probed by Southern hybridization. - [Read Mapping Dnase-I-hypersensitive Sites Protocol]
Dnase I is used to fragment a radiolabeled target DNA in the presence and absence of a nuclear extract. A "footprint" is generated when a protein binds to the target and protects a specific segment of DNA from the nucleolytic activity of Dnase I. By comparing the electrophoretic mobility of the Dnase I cleavage products to those of a sequence ladder derived from the same DNA fragment, the position(s) of the DNA sequences recognized by DNA-binding proteins can be determined. - [Read Mapping Protein-binding Sites on DNA by Dnase I Footprinting Protocol]
This protocol describes a method for reverse transcriptase (RT) in situ PCR. In situ PCR differs from PCR in situ hybridization in the inclusion of a reporter molecule in the amplification step. The two steps of RT in situ PCR that differ from in situ PCR are overnight digestion in RNase-free DNase that is performed after protease digestion, and an RT step, prior to in situ PCR. - [Read Reverse Transcriptase In Situ PCR Protocol]
Protocol describes a method for reverse transcriptase (RT) in situ PCR. In situ PCR differs from PCR in situ hybridization in the inclusion of a reporter molecule in the amplification step. The two steps of RT in situ PCR that differ from in situ PCR are overnight digestion in Rnase-free Dnase that is performed after protease digestion, and an RT step, prior to in situ PCR. - [Read Reverse Transcriptase In Situ PCR Protocol]
The key preparatory steps. In-situ RT-PCR. Fixative. Protease Digestion. DNase digestion. One-step in-situ RT PCR Protocol. Gerard J. Nuovo - [Read RT In Situ PCR - Foundation of Success]
Run-on transcription monitors the regulation of transcription in isolated nuclei. Protocol includes: Preparation of Nuclei, Run-On Transcription Reaction, Alkaline Hydrolysis of Run-Off Transcripts, Preparation of Filters and Hybridization. Solutions included: DNase Solution, Hybridization Mix, Denhardt Solution (100X) etc. - [Read Run- On Transcription Protocol]