Safety information - Eurogentec
Safety information - Eurogentec
Safety information - Eurogentec
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18<br />
The Tm in the dissociation curve for a specific miRNA is different<br />
from one sample to another<br />
Variations in extraction methods<br />
Differences in Tm can be explained by differences in pH, salt or detergents in a<br />
reaction due to variations in different extractions.<br />
Modification of the same miRNA from one cell type to another<br />
It could be possible that a product of different size or small modifications of the same<br />
miRNA are obtained from one cell type to another. To determine which miRNA is<br />
amplified, PCR product should be cloned and sequenced.<br />
MiRNA Editing<br />
Different mechanism of maturation of pri-miRNA to pre-miRNA (miRNA editing) can<br />
occur from one cell type to another. To determine which miRNA is amplified, PCR<br />
product should be cloned and sequenced.<br />
Specificity of MirMaid miRNA set to precursor miRNA versus<br />
primary miRNA<br />
Sequences of pre-miRNAs are identical to sequences of pri-miRNAs. Then, the<br />
MiRmaid qRT-PCR primer set will amplify both the pri-miRNA and the pre-miRNA.<br />
Primary miRNAs are 10 to 100 times less abundant than pre-miRNAs. Thus, the<br />
amplification would be mainly reflecting the quantification of the precusor miRNAs.<br />
To obtain a precise quantification of the pre-miRNAs versus the pri-miRNAs, specific<br />
primers to pri-miRNAs can be designed and quantification of primary miRNAs can be<br />
subtracted; thus, precise expression rate of miRNA precursors will be obtained.<br />
Manual for MiRmaid miRNA precursor qRT-PCR Primer Set I www.eurogentec.com I info@eurogentec.com
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