ChIP-seq of Tomato Fruit
1. Crosslinking: Treat fresh tomato greenripe fruit tissue with 1% formaldehyde to crosslink DNA and associated proteins in vivo. 2. Chromatin Isolation & Sonication: Extract chromatin and fragment it to 200500 bp via ultrasonic shearing. 3. Immunoprecipitation: Incubate fragmented chromatin with a specific antibody to enrich target proteinDNA complexes. 4. Decrosslinking & DNA Purification: Reverse crosslinks at high temperature, then purify the enriched DNA fragments. 5. End Repair & Adapter Ligation: Repair DNA ends, add Atails, and ligate sequencing adapters. 6. PCR Amplification: Amplify the ligated DNA to construct the ChIPseq library. 7. Quality Control: Verify library size and concentration using Agilent 2100 and Qubit.
SUB16121290
AJCHS2240909066IN