Method and kit for purifying nucleic acids
A nucleic acid and filter technology, used in the field of separation and/or purification of nucleic acids
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Embodiment 1
[0107] Example 1: Automated RNA Extraction from Nasopharyngeal Aspirates
[0108] Combined with large-porosity Erikoni that fit into 1.2ml Eppendorf pipette tips Matrix, 2ml deep well plate (USAScientific), Iconic Extraction reagents and nasopharyngeal aspirate as sample matrix were performed using an Eppendorf epMotion5070 liquid handling robot. The epMotion5070 liquid handling robot only holds no more than 8 tips simultaneously, so the baseline automation procedure is described for 8 parallel extractions. However, up to 24 samples can be processed during a single program in one deep well 96-well sample plate. For processing 16 or 24 samples, a separate epMotion program is available (and required). The procedure outlined below was used for the 8-sample automation script.
[0109] set up:
[0110] 1.1 Bring the nasopharyngeal sample to room temperature and start the extraction.
[0111] 1.2. Add an aliquot of 100 μL nasopharyngeal aspirate plus 150 μL nuclease-free...
Embodiment 2
[0144] Embodiment 2: the automated extraction of genomic DNA
[0145] A Hamilton STAR liquid handling robot was used to demonstrate the automated extraction of 96 samples from whole blood simultaneously. The Hamilton STAR differs from the epMotion system by having an optional heater / shaker unit on the panel, which is important for enzymatic digestion of certain clinical matrices such as whole blood. Because the system can be fitted with 96-channel pipette heads, there are dedicated 96-well plates for each of the filter tip steps and reagents.
[0146] set up:
[0147] 2.1 Turn on the STAR instrument and computer.
[0148] 2.2 Open the Hamilton operation control software.
[0149] 2.3 Open the run file provided by Icony for 96 samples.
[0150] 2.4 Place the labware on the STAR panel, such as Figure 5 shown.
[0151] 2.5 Distribute the reagents into their corresponding tanks according to Table 3 (the volume represents the minimum amount required to process 96 samples...
Embodiment 3
[0190] Example 3: Procedures for Purification of Fetal Nucleic Acids
[0191] Non-invasive prenatal diagnosis (NIPD) is an important and fast-growing market offering breakthrough medical advances due to its ability to replace standard prenatal diagnostic methods with many risks including fetal malformation and miscarriage. In contrast, tests for genetic abnormalities in fetal DNA present in the mother's plasma require only a simple blood draw. While this approach offers a lower-risk approach for prenatal diagnosis, there are challenges with sample types that require special handling techniques. First, fetal DNA is present in low concentrations in maternal plasma early in pregnancy, so the ability to handle large sample volumes and concentrate them in sufficient quantities for analysis is important. However, kits currently available on the market only allow input volumes of 250 μl to 5 ml and isolation of total nucleic acid. Second, fetal circulating DNA is present in mater...
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