Table 1 Comparison of LSplex labelling methods
From: Large scale multiplex PCR improves pathogen detection by DNA microarrays
Labelling Method | Description | Final amount of DNA1 (ÎĽg) | Base/Dye ratio2 | Labelled nucleotides | Processing time |
|---|---|---|---|---|---|
Random Priming | labelling after amplification with Klenow DNA polymerase | 1.3 | 61 | dCTP-Cy3 | 1.5 h LSplex, 15 min purification; 2 h labelling, 15 min purification |
Chromatide | direct incorporation of fluorescent nucleotides during Lsplex | 0.7 | 139 | Alexa Fluor 546-14-dUTP(1:3)3 | 1.5 h LSplex, 15 min purification |
ARES | incorporation of amino-modified nucleotides during Lsplex staining with Amino-reactive dye | 1.1 | 64 | aminoallyl-dUTP (1:2)4 stained after PCR with Alexa Fluor 555 | 1.5 h Lsplex, 15 min purification; 1 h post staining, 15 min purification |