The Bisulfite Primer Seeker is a free bisulfite primer design tool that generates primers for Bisulfite Specific PCR (BSP). This free bisulfite primer design tool simplifies the tedious process of bisulfite primer design. Successful bisulfite primer design is critical to unbiased, region-specific DNA...
TaKaRa EpiTaq HS (for bisulfite-treated DNA) 说明书
Custom design primers EpiMark Hot Start Taq DNA Polymerase (NEB #M0490) or other hot start DNA polymerase specific for bisulfite converted DNA. Heat block or water bath (requiring temperatures of 65°C and 92°C) 96–100% molecular biology grade ethanol PCR Thermal Cycler 0.2 ml strip...
amplificationandcloningofPCRproductsamplifiedfrombisulfite-convertedDNA samples.Optionalprotocolsdetailapromisingrapidbisulfitetreatmenttechnique(Shiraishi andHayatsuetal.,2004)andgenomicDNApreparationfromfixedtissue(Aietal.,2008), greatlyextendingpotentialapplicationsofBGS.Asupportingprotocolcoversproperprimer ...
(Clarket al.,1994). Methylcytosine may then be detected by standard DNA sequencing of the PCR products (Figure 14). The most important aspect of sequencing bisulfite-modified DNA is the primers design. Three factors need to be considered in the design of PCR primers for sequencing bisulfite-...
First, your samples are checked to confirm the quality and concentration and then converted using theEZ DNA Methylation-Lightning Kit. The flux capacitor is then set for maximum travel speeds (optional and highly variable). Next, post-bisulfite PCR primers for each target region are designed usin...
s protocol. DNA strands containing a specific sequence tag from random primers were synthesized. Then, a known sequence tag was added to the 3′-end of DNA strands. The di-tagged DNA was purified by using 1.6X AMPure XP beads. The library was amplified by the Failsafe PCR enzyme system...
The 19.5 μl eluate was placed in a 0.2 ml PCR tube preloaded with a PCR reaction mixture containing 25 μl of 2x KAPA HiFi HotStart Uracil+ ReadyMix (Roche, KK2801), 1.5 μl of 25 μM P5 transposon primer (ordered from IDT with the sequence shown in Supplementary Fig...
Methylation-specific PCRObjectives To verify complete bisulfite modification. Design and methods Bisulfite modified DNA was screened by PCR with Calponin-specific primer sets prior to detecting methylation status of 14-3-3σ by methylation-specific PCR. Results False positive methylation was obtained from...
Quantitative real time PCR 500 ng of total RNA was reverse transcribed using an oligo(dT)12 to 18 primer with Superscript II reverse transcriptase (Invitrogen), according to the manufacturer’s instructions. Reverse transcription PCR primers were designed between different exons to avoid any amplifica...