DNA重组

IntroductionIncreasing evidence in literature reveals that the organization of chromosomal domains in the cell nucleus plays an important role in gene expression during cellular differentiation and de ...

IntroductionSequencing of sodium-bisulfite modified genomic DNA originally introduced by M. Frommer (Frommer et al. 1992) is a widely used “gold standard” method for DNA-methylation analysis. Since th ...

Introduction(based on Olek et al. 1996 Schoenherr et al. 2003)DNA methylation is a stable epigenetic mark which can mediate gene silencing. Bisulfite sequencing allows for precise identification of me ...

IntroductionGenomic DNA methylation is one of the most important epigenetic modifications in eukaryotes. It is essential for life and its alteration is often associated with disease. In animals most o ...

IntroductionMethylation of cytosines can mediate epigenetic gene silencing and is the most prominent DNA modification in eukaryotes. MeDIP is an immunocapturing approach to enrich DNA that is methylat ...

IntroductionNucleosomal positioning plays a pivotal role in the regulation of transcriptional initiation. Transcriptional co-activator complexes interact with nucleosomes (4) to induce nucleosomal re ...

IntroductionThe following "bisulfite genomic sequencing" protocol has been optimized for the determination of DNA cytosine methylation at transgene loci in plants. The conditions used are based on the ...

IntroductionEfficient duplication of eukaryotic genomes relies on the sequential activation of thousands of replication origins distributed along the chromosomes. This process follows a complex spatia ...

Collect fresh tissue and store at -20°C. Grind leaf tissue to a fine powder with liquid nitrogen in a prechilled mortar . Transfer ground tissue with a chilled paintbrush into a well-chilled 50-ml pol ...

DNA Digestion Make a cocktail as follows: Southern hybridization cocktail 10 X buffer3.50 µLBSA 0.35 µLddH205.65 µLRNase 0.50 mLEcoRI 10.00 µLTOTAL20.00 µL/tubeAdd 15 μL DNA to each tube containing 2 ...

Prehybridization Set hybridization oven to 65°C. Thaw 32P isotope in hood. Place membrane in hybridization tube. Add 50 mL of prehybridization buffer to each tube. Prehybridization buffer ddH20125 mL ...

Hybridization Hybridization buffer ddH2025.0 mL20X SSPE12.0 mL100X Denhardt's solution2.0 mL20% SDS1.0 mLSalmon sperm DNA 0.2 mLdextran sulfate2.0 gTOTAL40.2 mLReplace prehybridization buffer in each ...

New D14 primers.Lr21L:CGC TTT TAC CGA GAT TGG TC Lr21R:TCT GGT ATC TCA CGA AGC CTT Lr21: 669 bplr21-Fielder: 757 bplr21-Wichita: 774 bpRecipe (in 25 µl reaction). Buffer (10X)2.5 µl2.5 mM dNTP2.5 µl25 ...

New D14 primers.Lr21L:CGC TTT TAC CGA GAT TGG TC Lr21R:TCT GGT ATC TCA CGA AGC CTT Lr21: 669 bplr21-Fielder: 757 bplr21-Wichita: 774 bpRecipe (in 25 µl reaction). Buffer (10X)2.5 µl2.5 mM dNTP2.5 µl25 ...

Developed for microarray-based comparative genomic hybridization.Genomic DNA can be labeled with a simple random-priming protocol based on Gibco/BRL's Bioprime DNA Labeling kit though nick translation ...

PrincipleSingle-strand conformation polymorphism (SSCP) technique is a simple and efficient means to detect any small alteration in PCR-amplified products. It is based on the assumption that subtle nu ...

Genomic DNA will be randomly primed with a sequence tagged oligonucleotide for 2 cycles. This will create random genomic products with a specific tag at both ends. These products will then be amplifi ...

Genomic DNA will be randomly primed with a sequence tagged oligonucleotide for 2 cycles. This will create random genomic products with a specific tag at both ends. These products will then be amplifi ...

The major considerations in using alcohol to precipitate DNA are:Temperature: -20°C is optimal but 0°C can be used for 20 ng/mL2) Amount: For small amount of DNA ( i.e. too little to reliably see a pe ...

Reagents:Lysis Buffer: (10 mM Tris pH8 100 mM NaCl 25 mM EDTA 0.5%SDS) store at room temp.10 mg/mL Proteinase K: 100 mg (Roche) + 10 mL buffer (10 mM Tris pH8.0 20 mM CaCl2 50% (v/v) glycerol) store ...