Methylation-specific PCR (MSP) is still the method of choice for a single gene methylation study. The proper design of the primer pairs is a prerequisite for obtaining reliable PCR results. Despite numerous protocols describing the rules for MSP primer design, none of them provide a comprehensive...
PCR primers can be designed with pen and paper, but—especially with the added concerns of MSP—“it just becomes inherently easier to do that with software,” says Herman, whose lab developed an application, MSPprimer, specifically to design primers for MSP [2]. MSPprimer incorporates all ...
Step 2: Primer Design PCR primer design for MS-HRM is critical—the free online tool from Applied Biosystems, Methyl Primer Express Software v1.0, was developed specifically for primer design in methylation studies (www.app...
DNAmethylationisanepigeneticmechanismofgeneregulation.Bisulfite-conversion-basedPCRmeth-ods,suchasbisulfitesequencingPCR(BSP)andmethyla-tionspecificPCR(MSP),remainthemostcommonlyusedtechniquesformethylationmapping.Existingprimerde-signprogramsdevelopedforstandardPCRcannothan-dleprimerdesignforbisulfite-...
B. Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands. Proc. Natl. Acad. Sci. USA, 93: 9821-9826, 1996. 19. Vo,... J Herman - 《Current Topics in Microbiology & Immunology》 被引量: 370发表: 2000年 Methylation specific PCR, PNAS 1996.pdf B. Methylati...
Methylation-specific PCR (MS-PCR or MSP) is one of the most commonly used methods for gene/sequence-specific detection of DNA methylation. The DNA undergoes bisulfite conversion of cytosine to uracil and then the methylated sequences are selectively amplified with primers specific for methylation. ...
Prediction of CpG islands in TP73 promoter and primer design for methylation-specific PCR was possible through the use of a web software (www.urogene.org); Primer pairs used for HepCY, HepCO, HepG2, SNU449, Caco2, HCT116 and HS27 cell lines in methylation-specific PCR at TP73 promoter...
then used for PCR amplification, adapter/index ligation, and DNA purification with a TruePrep® DNA Library Prep Kit V2 for Illumina (Vazyme, Cat.# TD502). BS-seq libraries were constructed using a previously reported protocol24with modified primer adapter 2 oligos and iPCRtag primers20. RNA...
High-yield, converted DNA is ideal for PCR, Methylation Specific PCR (MSP), arrays, library preps, Next-Generation sequencing, etc. DESCRIPTION The EZ-96 DNA Methylation-Lightning MagPrep Kit is the fastest magnetic bead-based DNA bisulfite conversion kit for high-throughput and automated methylati...
For indexing PCR, the following was added to each well of the ligation plate: 10 µL 5X VeraSeq GC Buffer, 2 µL 10 µM dNTPs, 1.5 µL VeraSeq Ultra Enzyme, 24 µL dH2O, 0.5 µL 100X EvaGreen, 1 µL 10 µM TruSeq i5 primers, 1 µL 10...