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| The principle of LAMP method |
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SNPs typing - LAMP-based SNPs typing |
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Because
of the high specificity of the LAMP method, only the target gene
will be amplified from gene samples containing homologous nucleotide
sequences when using LAMP-based SNPs typing. Furthermore, because
of the characteristics of its amplification reaction (c.f. basic
principle), the LAMP method discriminates a single nucleotide difference
at each cycling step of the DNA replication, through both "sense
and anti-sense strand" reactions, and the type of SNP can easily
be detected just by amplifying the DNA containing SNP in a single
step.
Due to the simplicity and rapidity of the LAMP method, simple detection
of SNPs typing can be achieved within 30 minutes. |
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With the use of 4
primers designed to recognize 6 distinct
regions, only the target gene is strictly
and specifically amplified even in coexistence
with its homologous gene. |
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The reaction is so
specific as to strictly discriminate single
nucleotide difference. |
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| The SNPs
typing can be achieved by detecting the presence
of amplified products in a single step. |
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The
figures below show the basic principles of the process when using
Wild Type (WT) Primers.
The FIP and BIP are designed to contain a SNP nucleotide (in this
case of Wild Type allele) at 5' end, respectively.
Using the WT primers, when the target gene is the WT allele, DNA
synthesis from dumbbell-like starting structure proceeds and the
LAMP amplification cycling continues. In contrast, when the target
gene is the Mutant (MUT) allele, no DNA synthesis proceeds from
dumbbell-like structure and the LAMP amplification cycling dose
not occur. Even if DNA synthesis proceeds in one step due to miscopy,
the amplification reaction is either halted in other steps or is
delayed since repetition of this reaction continually checks at
each cycling step of the DNA replication.
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