A Polymerase That Makes the Most out of Every NGS Experiment
NGS applications like long-range multiplex PCR for targeted resequencing require high-quality amplicons. Choosing the right polymerase for library prep promotes even coverage and successful sequencing.
PrimeSTAR GXL DNA Polymerase is known for its:
- Surefire amplification of GC- or AT-rich, long, and repetitive templates
- High specificity due to hot-start technology and pre-optimized buffer
- Highest processivity of all commercially available high-fidelity DNA polymerases
Excellent product. We PCRed the long fragment that we failed to get with other polymerases.
Excels in Multiplex PCR for Genotyping Applications
[Our multiplex PCR-NGS] method for the nine HLA loci greatly simplifies the procedures required in preparing the DNA samples for NGS by reducing the time of preparation and the amount and costs of reagents, including the use of much smaller amounts of template DNA samples.
Electrophoresis images of PCR products from two unrelated genomic DNA samples. In Figure 4 from their paper, the authors used PrimeSTAR GXL DNA Polymerase to amplify long-range PCR products for template DNA amounts ranging from 1 ng to 25 ng. The lanes labeled M are bands of the 1 kb DNA size marker ladder. DNA yield after purification of the PCR products ranged from 119.7 ng (1 ng template) to 608.6 ng (25 ng template) in Sample 1, and from 112.5 ng (1 ng template) to 559.7 ng (25 ng template) in Sample 2. 100 ng of the multiplex PCR products were used for the preparation of each Ion PGM library. Sequence reads had high quality and sufficient sequence volume for further HLA genotyping analysis. Figure is modified from figure 4 of Ozaki, et al., 2015 under Creative Commons License 4.0.
Supports Even Sequence Coverage
We found that… PrimeSTAR GXL DNA polymerase can amplify almost all amplicons with different sizes and Tm values under identical PCR conditions. Other enzymes require alteration of PCR conditions to obtain optimal performance.
Visualization of sequencing read depth in SeqMonk for long-range amplicon BRCA1.6. The authors chose PrimeSTAR GXL DNA Polymerase over five competing enzymes to prepare libraries for sequencing using the Illumina® MiSeq® platform. Figure 4 from their paper shows analysis of three amplicons—BRCA1.1, BRCA1.6, and BRCA2.8—and selected data are shown here. Although variations of read depth exist within and between amplicons from long-range PCR, good coverage of all amplicons was observed. Here, coverage is Log2 transformed; the right column shows a junction region. Figure is modified from Figure 4 of Jia, et al., 2014 under Creative Commons License 4.0. For more about this study, please view our Case Study, “Long PCR for Cost-Effective, Efficient Targeted Next-Gen Sequencing.”