| Autor: |
Mattocks, Chris, Harvey, J.F., Cross, N.C.P. |
| Předmět: |
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| Zdroj: |
Journal of Medical Genetics; Sep2003 Supplement, Vol. 40, pS78-S78, 1/4p |
| Abstrakt: |
The AutoGen NA-3000EU automatic nucleic acid isolation system is a fully automated instrument-chemistry designed for the isolation of genomic DNA and RNA from various animal tissues. The robot is preprogrammed with eight standard protocols that can be modified by the user as required and is capable of processing 48 samples in a single run. We have carried out a detailed evaluation the two DNA extraction protocols considered to be most applicable to diagnostic laboratories: (i) A low volume protocol (0.05-1.0 mi whole blood) and (ii) a high volume protocol (up 7 mi whole blood). We prospectively extracted 200 samples for each protocol and analysed the resulting DNAs for yield, purity and PCR efficiency. We have also carried out comparative studies on >50 samples with respect to a manual salt extraction protocol by splitting samples. In the initial study there was a failure rate of 15% for the automated protocol compared to 0% for the manual method. DNA yield was found to be approximately half that of the manual method (26 ug/ml blood [auto], 56 ug/ml blood [manual]). Purity was good with respect to protein (average A260/A 280:1.77 [auto], 1.89 [manual]) but was more variable than with the manual method (SD: 0.22 [auto], 0.04 [manual]). Some Isopropanol contamination was detected, although this did not appear to affect the efficacy in PCR, which was equivalent to that for manually extracted DNA. Further work to reduce the failure rate and increase yield is ongoing and will be presented together with costing and evaluation of the usability of the system. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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