GC-RICH DNA AMPLIFICATION

GC-rich DNA Amplification

Amplification of DNA with a high percentage of GC content is often compromised by inadequate strand separation and complex secondary structure formations. The inability of the DNA polymerase to read through strong secondary structures such as hairpins, results in very low PCR yield and formation of truncated PCR products.

Ampliqon has developed a unique solution for amplification of GC-rich and other difficult DNA targets.

GC Buffer I or GC Buffer II in combination with TEMPase Hot Start DNA polymerase promotes excellent amplification results of DNA targets with varying high degrees of GC content.

  • GC-rich DNA Target Kit and GC TEMPase master mixes
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GC TEMPase master mixes, GC Buffer I and GC Buffer II are manufactured in-house in accordance to our ISO 9001:2015 quality management system, ensuring that each batch possess the same robust performance every time.

GC-rich DNA Target Kit

GC-rich DNA Target kit is specially developed for the amplification of GC rich DNA sequences.

GC-rich DNA Target Kit offers considerable flexibility and can be used as an initial testing kit before you use one of our GC TEMPase 2x master mixes.


FEATURES

  • High success rate with amplification of GC-rich DNA
  • High sensitivity, specificity and product yield
  • Diminished formation of non-specific products
  • Reaction set-up at room temperature

DESCRIPTION

The GC-rich DNA Target Kit consists of TEMPase Hot Start DNA Polymerase, 4x GC Buffer I, 4x GC Buffer II and MgCl2, all in separate tubes. MgCl2 is enclosed in the GC-rich DNA Target Kit, to ensure convenient optimisation. 

4x GC Buffer I and II are also available separately.

TIP - WHEN TO CHOOSE SPECIALISED GC BUFFERS

If your PCR fails with TEMPase Hot Start Polymerase in combination with Ammonium Buffer, we kindly recommend you to try TEMPase in combination with GC Buffer I either as master mix or GC-rich DNA Target Kit.  If your amplification is still not satisfactory, then switch to GC Buffer II.

To save time both GC buffers can be tested at the same time.

OPTIMISATION OF GC-RICH DNA AMPLIFICATION

Six genes with varying percentage of GC contents were amplified with Standard Buffer (lanes S), Ammonium Buffer (lanes A), GC Buffer I (lanes I) and GC Buffer II (lanes II). M. Marker. With an increasing percentage of GC content in the expected amplicon, Standard Buffer and Ammonium Buffer fail to support the correct amplification products, while GC Buffer I and GC Buffer II succeed. Correct amplified products are encircled.

PRODUCT INFO

GC-rich DNA Target Kit

With GC Buffer I, GC Buffer II and MgCl2
4x GC Buffer I
4x GC Buffer II
Sample

GC TEMPase master mixes

GC 2x master mixes are ready-to-use mixes based on either GC Buffer I or GC Buffer II. GC TEMPase master mixes are specifically developed for the amplification of GC-rich DNA sequences.


FEATURES

  • High success rate with amplification of GC-rich DNA
  • High sensitivity, specificity and product yield
  • Diminished formation of non-specific products
  • Reaction set-up at room temperature

DESCRIPTION

GC-rich 2x master mixes consist of TEMPase Hot Start DNA Polymerase combined with either GC Buffer I or GC Buffer II, dNTPs and MgCl2. Just add DNA template and primers to successfully carry out PCR.

4x GC Buffer I and II are also available separately.

TIP - WHEN TO CHOOSE SPECIALISED GC BUFFERS

If your PCR fails with TEMPase Hot Start Polymerase in combination with Ammonium Buffer, we kindly recommend you to try TEMPase in combination with GC Buffer I either as master mix or GC-rich DNA Target Kit.  If your amplification is still not satisfactory, then switch to GC Buffer II.

To save time both GC buffers can be tested at the same time.

OPTIMISATION OF GC-RICH DNA AMPLIFICATION

Six genes with varying percentage of GC contents were amplified with Standard Buffer (lanes S), Ammonium Buffer (lanes A), GC Buffer I (lanes I) and GC Buffer II (lanes II). M. Marker. With an increasing percentage of GC content in the expected amplicon, Standard Buffer and Ammonium Buffer fail to support the correct amplification products, while GC Buffer I and GC Buffer II succeed. Correct amplified products are encircled.

COVERAGE STUDY OF GC TEMPASE MASTER MIX I

GC TEMPase I displays superior performance on a broad range of amplicons with a GC content ranging from 41 – 78 %. Five genes with increasing percentage of GC content were amplified using GC TEMPase Master Mix I. Lane M is Marker (Iqon DNA Mini Ladder). Number of PCR cycles used in this study is 40

PRODUCT INFO

GC TEMPase master mixes

2x Master Mix I
Sample
2x Master Mix II
Sample