Epigenomics Profiling DNA Methylation Reduced 50 mg Representation Bisulfite Sequencing

Introduction

Methylation is an important method of modifying a nucleic acid. The result of a methylated nucleotide in the right position will regulate the gene’s expression. For this reason, methylation is one of the most important aspects contributing to epigenetics and is closely related to many diseases, such as cancer, aging, and Alzheimer. CELLFREE can provide DNA methylation, RNA methylation and related products to our customers.

Services

• Bisulfite modification and sequencing method (BSP)

Frommer et al (1992) proposed methods of DNA methylation analysis, this method is reliable and precise. Procedures:

1. Prepare genomic DNA of Sample

2. Bisulfite treatment (including purification and de-sulfosalicylic reaction) to convert unmethylated cytosine to uracil, while methylated cytosine remains unchanged.

3. Using methylation primer design software to design primers at the both ends of methylated regions.

4. Fragment amplification and PCR product recovery.

5. PCR products were sequenced directly or after cloned (quantitative analysis), analysis of each CpG island methylation.

6. Data Statistics. unmethylated DNA samples are used as a control.

• Methylation specific PCR method (Methylation-Specific PCR, MS-PCR)

Herman et al (1996) in the use of bisulfite treatment on the basis of the new methods. High sensitivity of this method is mainly used for qualitative research. Procedures:

1. Prepare genomic DNA of Sample

2. Bisulfite treatment (including the purification and de-sulfosalicylic reaction).

3. Using methylation primer design software design methylation specific primers (primer 1) or non-methylated primers in methylated regions (primer 2) 2 pairs.

4. For specific PCR amplification, only the fragments fully combinate with methylated or non-methylation specific primers can amplify the product.

5. Detection of MSP amplification, if DNA methylation for the treatment chain primers amplified fragments, it shows that the detected methylation sites exist; if used for the treatment of non-methylated DNA chain The primers amplified fragments, then the detected methylation sites does not exist.

Customer offer

Fresh and sufficient sample (tissue not less than 50 mg, whole blood not less than 500 μl), or purified DNA (≥ 5 μg/sample), genetic information detected , related literature, etc.

We provide

Experimental report, MSP electropherogram / BSP sequencing map.