LA Taq DNA Polymerase with GC Buffers
LA Taq DNA Polymerase with GC Buffers
Brand: Takara Bio.
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LA Taq DNA Polymerase with GC Buffers
LA Taq DNA Polymerase with GC buffers
TaKaRa LA Taq DNA Polymerase with GC Buffer has the TaKaRa LA Taq DNA Polymerase blend for long-and-accurate PCR paired with GC-optimized buffers to power through difficult-to-amplify GC-rich sequences, including genomic DNA. The GC-optimized Buffers I and II are specifically designed to amplify GC-rich DNA templates (up to 73% GC content) or templates with a significant amount of secondary structure:
- GC Buffer I is recommended for amplification of fragments that are ≥5 kb in length
- GC Buffer II is recommended for fragments that are 2–3 kb in length
For the longest amplicons that are also GC rich, try PrimeSTAR GXL DNA Polymerase, which is designed to amplify genomic DNA up to 30 kb.
Overview
- Long and accurate PCR amplification of genomic DNA targets (long-range PCR)
- Optimized buffers (GC Buffers I & II) for amplification of templates with high GC content and/or significant secondary structure
- High-yield amplification of long DNA targets with minimal optimization
PCR products
PCR products generated with Takara LA Taq contain a mixture of 3'-A overhangs and blunt ends, which allows >80% cloning efficiency into T-vectors. However, the efficiency of cloning longer products (>5 kb) into T-vectors is quite low. It is also possible to clone the product in blunt-end vectors after blunting and phosphorylation of the end.
Comparison of Takara LA Taq with GC Buffer and Takara LA Taq DNA polymerase for amplification of a GC-rich target fragment
Comparison of Takara LA Taq with GC Buffer and Takara LA Taq DNA polymerase for amplification of a GC-rich target fragment. A 2.1 kb mouse rRNA gene (63.4% GC) was amplified from of mouse genomic DNA. Takara LA Taq with GC Buffer resulted in high yield, high specificity amplification.
Lanes:
M1: lambda-Hind III digest
1 and 2: LA Taq / LA PCR Buffer II
3 and 4: LA Taq with GC Buffer / GC Buffer I
5 and 6: LA Taq with GC Buffer / GC Buffer II
M2: pHY Marker
Amplification of the Huntington's Disease (HD) gene using Takara LA Taq DNA Polymerase
Amplification of the Huntington's Disease (HD) gene using Takara LA Taq DNA Polymerase. The HD gene is GC-rich and highly repetitive and the primers used in this experiment amplified regions of the HD gene IT15 CAG repeat. For all reactions, purified human genomic DNA (100 ng in a 50 µL reaction) was used as a template in reactions containing Takara LA Taq polymerase with either LA PCR Buffer II (lane 1), GC Buffer I (lane 2), or GC Buffer II (lane 3), or a competitor's high GC DNA polymerase kit (lane 4). The sizes of the amplified products were 262 bp (GC content 73%) and 358 bp (GC content 71.5%). Lane M contains a 100 bp ladder.
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Comparison of Takara LA Taq with GC Buffer and Takara LA Taq DNA polymerase for amplification of a GC-rich target fragment
Comparison of Takara LA Taq with GC Buffer and Takara LA Taq DNA polymerase for amplification of a GC-rich target fragment. A 2.1 kb mouse rRNA gene (63.4% GC) was amplified from of mouse genomic DNA. Takara LA Taq with GC Buffer resulted in high yield, high specificity amplification.
Lanes:
M1: lambda-Hind III digest
1 and 2: LA Taq / LA PCR Buffer II
3 and 4: LA Taq with GC Buffer / GC Buffer I
5 and 6: LA Taq with GC Buffer / GC Buffer II
M2: pHY Marker
Amplification of the Huntington's Disease (HD) gene using Takara LA Taq DNA Polymerase
Amplification of the Huntington's Disease (HD) gene using Takara LA Taq DNA Polymerase. The HD gene is GC-rich and highly repetitive and the primers used in this experiment amplified regions of the HD gene IT15 CAG repeat. For all reactions, purified human genomic DNA (100 ng in a 50 µL reaction) was used as a template in reactions containing Takara LA Taq polymerase with either LA PCR Buffer II (lane 1), GC Buffer I (lane 2), or GC Buffer II (lane 3), or a competitor's high GC DNA polymerase kit (lane 4). The sizes of the amplified products were 262 bp (GC content 73%) and 358 bp (GC content 71.5%). Lane M contains a 100 bp ladder.
