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Regenerates and
amplifies damaged DNA template
- Optimized
for PCR
- Optimized
enzyme blend for PCR with simultaneous damaged
template regeneration
- Greatly
improves chances of success when amplifying
poor-quality template
- Adapted
to amplifications from 500 bp to 11 kb
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Reference Library
Product
profile |
Standard thermostable polymerases are unable to repair damaged
template containing, for example, DNA strand breaks and base
lesions. Degradation of DNA template can occur for numerous
reasons: physical shearing during extraction or purification,
successive freeze-thaw steps, non-optimum storage, aging,
depurination at low pH, UV exposure and irreversible
denaturation, etc. Template damage of this kind is unavoidable
with, for example, forensic samples, some specimen preservation
methods or certain aggressive methods of DNA extraction.
The
Auroris™ PCR Repair Kit is recommended for use in any PCR
where degradation is suspected or where extra specificity and
sensitivity is required. This kit can regenerate most damaged
templates, depending on the level of degradation, and provides a
specific PCR product.
This kit
comprises two special polymerase formulations :
MIX Pol A : Used for amplifying degraded fragments over 4
kb in length (up to 11 kb using genomic DNA and up to 20 kb using
phage DNA)
MIX Pol B : Used for amplifying degraded fragments from
500 bp to 7 kb.
The Auroris™ PCR Repair Kit has been developed in
collaboration with INSERM (Institut National de la Sante et de la
Recherche Medicale).
Note
: not suitable for amplification of sequences < 500 bp
Figure
1 :
Amplification of a 900 bp fragment from a human -globin gene.
D1: non degraded DNA template
D2: denatured DNA at 99C for 60 sec
D3: denatured DNA at 99C for 90 sec
D4: denatured DNA at 99C for 120 sec
D5: DNA extracted from blood cells found thawed after a period of
2 months
Taq DNA Pol (P1), Arrow Taq DNA Pol (P2), Mix Pol A (A) and Mix
Pol B (B) were compared. 1 U of DNA Pol, 1 ng DNA template, 100 M of each dNTP and 50 pmoles of specific primers were used.
PCR program: (5 min at 93C) x 1 . (1 min at 91C, 1 min at
62C, 1 min 30 sec at 72C) x 35
Storage
buffer: 20 mM TrisHCl, pH 8.0, 100 mM KCl, 0.1 mM EDTA, 1 mM
Dithiothreitol, 0.5%Tween 20, 0.5% Nonidet P40, 50% Glycerol
Incubation conditions: 10 mM TrisHCl, pH 9.0, 50 mM KCl,
1.5 mM MgCl2, 0.1% Triton X100, 0.2 mg/ml BSA
Quality control: Activity, absence of nickases and
endonucleases, specific PCR on altered and non altered genomic
and phage templates.
Unit definition: One unit is the amount of enzyme required
to catalyse the incorporation of 10 nmol of nucleosides into acid
soluble material in 30 min at 74C under assay conditions.
| EPAPK050 |
Auroris™
PCR Repair Kit |
50U of each
Mix Pol (at 1U/l) |
| EPAUS050 |
Auroris™ Mix Pol A |
50U at 1U/µl |
| EPBOR050 |
Auroris™ Mix Pol B |
50U at 1U/µl |
Storage:
-20C
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