HaeIII Methyltransferase

PCR Methylation specific PCR - Bacterial DNA

Experiment
PCR Methylation specific PCR - Bacterial DNA
Product
HaeIII Methyltransferase from New England BioLabs
Manufacturer
New England BioLabs

Protocol tips

Protocol tips
S-adenosylmethionine (SAM) is unstable at (pH 7.5), 37°C and should be replenished in reactions incubated longer than 4 hours.

Publication protocol

Analysis of methylation by M.CbeI
To identify the site of methylation by M.CbeI, unmodified DNA was compared to that after methylation and the changes were determined by direct visualization in automated DNA sequencing chromatograms , . methylation of pUC18 DNA isolated from DH5α () was carried out using M.HaeIII (NEB) and purified M.CbeI. The efficiency of methylation was determined by cleavage of the methylated and unmethylated DNA with HaeIII (NEB), purified CbeI, and cell free extracts (CFE). Digested DNA was displayed by agarose gel electrophoresis and visualization using ethidium bromide staining. Automatic sequencing was performed using primers M13F(−20) and M13R(−20) in an ABI automated PRISM big-dye-terminator system (Macrogen, Inc, Maryland). Sequences were analyzed using the Chromas Lite v2.01 (Technelysium Pty Ltd.) and ABI chromatograms were compared by aligning the Sequencing traces and using SeqDoc .

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Discussion

Discussion

5 years ago

Author: United Kingdom

How “strong” is the PCR product of methylation specific PCR?

Does anyone know how “strong” the PCR product of methylation specific PCR is? I kept my PCR products at 4C for about 3 weeks and then at room temperature for another week. Will I be able to use them for sequencing?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing PCR Methylation specific PCR - Bacterial DNA using HaeIII Methyltransferase from New England BioLabs.

Paper title
Methylation by a Unique α-class N4-Cytosine Methyltransferase Is Required for DNA Transformation of DSM6725
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Manufacturer protocol

Download the product protocol from New England BioLabs for HaeIII Methyltransferase below.

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