*HiFi-Polymerase is our self-purified DNA polymerase
10x HiFi buffer - to be used with any polymerase
200 mM Tris-HCl, pH 8.8
100 mM (NH₄)₂SO₄
500 mM KCl
1% (v/v) Triton X-100
1 mg/ml BSA
20 mM MgCl₂
We note that the Phusion polymerase using the manufacturer supplied buffer does not work for cassette amplification with M1 and M2 tagging oligos,
whereas for Velocity polymerase using the buffer provided by the manufacturer good amounts of the product are obtained.
We suggest pipetting on ice and pre-heating the PCR machine or hot start.
We found that all polymerases work well using the buffer conditions and amplification above.
The information contained in this website is correct to the best of our knowledge.
Under no circumstance shall the authors be liable for any potential mistakes, claim, damage or loss arising from the use of the application
'Online oligo design tool for PCR tagging in mammalian cells'.
The use of the tool and the reliance on any information on the site is solely at the user's own risk.
Only for non-commercial, academic or research purposes.
29.10.2019 Oligo ranking updated - cleavage after STOP codon preferred
22.05.2019 Template list completed with Addgene links to mNeonGreen plasmids