Shanghai methylation test experiment

Shanghai Methically Test Experiment

Service Project

[123 1, methylation-Specific PCR (MSP)

Treatment of genome DNA with bisulfite, and the methylated cytosine is converted to uracil. The methylated cytosine is unchanged; the primer for methylation and unoformat sequences are then designed to perform PCR. The MSP amplification product is detected by electrophoresis, and if the amplified fragment is obtained by the primer of the methylated DNA chain after treatment, the site is presented by methylation; in turn, it will be described that there is no methylation in the detected site.

2, Bisulfite Sequencing PCR, BSP)

Treatment of genomic DNA with bisulfite, the methylated cytosine is converted to uracil. The methylated cytosine is unchanged. The BSP primer was subsequently designed to perform PCR, and the uracil was transformed into thymine during amplification, and then the PCR product was sequenced to determine whether the CPG site was modified as a BSP-direct sequencing method. The PCR product was cloned to the carrier to be sequenced, and the sequencing success rate can be improved, which is called the BSP-clone sequencing method.

3, Methyleization-Sensitive Amplified Polymorphism, MSAP

MSAP is the use of two homozymes HPA II for DNA methylation. MSP I is to perform enzyme digestion and connections to genomic DNA. Since the two enzymes are capable of identifying the same restriction enzyme sink, the CCGG site. When the CCGG site in the DNA sequence has different degrees of methylation state, it is identified by these two enzymes, respectively, and is embodied in a non-product.

4, pyrosequencing

by accurate quantitative amount of methylation frequencies on a single continuous CPG site, pyrophosphate sequencing can detect and quantitatively quantitatively Change. During the sequence extension, the C-T ratio of a single bit point is determined according to the amount of incorporated in C and T. Therefore, methylation variation of different sites can be accurately detected. Due to the actual sequence data, the methylation state is presented in the sequence form.

Send sample requirements

Cell ( u0026 GE; 106), tissue ( u0026 GE; 300 mg), blood ( u0026 ge; 1ml), serum ( u0026 ge; 1.5ml) Sample materials, genomic DNA (body)Product u0026 GE; 20 u0026 Mu; L, concentration u0026 ge; 50 ng / u0026 mu; l).