Takara enzyme for GC-rich target sequences

Consider the following enzymes: 1. First choice: PrimeSTAR GXL DNA Polymerase is the most effective enzyme for GC-rich templates, such as bacterial genomic DNA. It facilitates high-fidelity amplifications with very few errors. PrimeSTAR GXL DNA Polymerase has been used successfully to amplify a region with ~70% GC content in a standard reaction using the buffer provided with the enzyme. 2. [...]

Takara enzyme with highest fidelity

PrimeSTAR polymerases provide better fidelity than Pfu DNA polymerase, which is widely considered to be a high-fidelity enzyme. PrimeSTAR Max DNA Polymerase has the highest fidelity; when this enzyme was used to amplify the entire pUC119 plasmid, sequence analysis detected only four mutations out of 370,656 total bases sequenced (an error rate of 0.0010%). In addition, compared to other enzymes, [...]

What precautions should be taken when using inosine-containing primers?

TaKaRa Taq DNA Polymerase and TaKaRa Taq DNA Polymerase Hot Start Version are compatible with inosine-containing primers. Inosine-containing primers should not be used with PCR enzymes that have 3'?5' exonuclease activity (e.g., PrimeSTAR HS DNA Polymerase, PrimeSTAR Max DNA Polymerase, PrimeSTAR GXL DNA Polymerase, TaKaRa Ex Taq DNA Polymerase, or Takara LA Taq DNA polymerases), nor with Terra [...]

PrimeSTAR Max and PrimeSTAR GXL DNA polymerases have very high fidelity; how was fidelity measured for these enzymes?

The fidelities of the PrimeSTAR polymerases were measured by Sanger sequencing of individual colonies after PCR, as described below: -Ten arbitrarily selected GC-rich regions of Thermus thermophilus HB8 genomic DNA were amplified. -PCR products were cloned into a plasmid vector. -Multiple clones were selected for each respective amplification product, and the PCR insert was sequenced. [...]

How can I compare error rates of different high-fidelity polymerases?

Error rates reported by vendors for polymerases cannot always be directly compared, as different methods are used to measure fidelity. These methods include: 1. Blue-white screening This approach is based on phenotypic changes and is widely used since it is fast, relatively simple, and cost effective. The original method for blue-white screening, known as the Kunkel method (Kunkel and Tindall [...]