Sample Preparation and Submission
In addition to isolated nucleic acids, amplicons and libraries MOgene has handled a wide variety of sample types – from microbial samples as cultures or cell pellets, insects, insect larvae, tissue samples from various vertebrate and non-vertebrate species and tissue samples from wide range of plant species for nucleic acid extractions.
DNA/ RNA EXTRACTION SAMPLES
Blood samples for DNA: 8.5 mL of whole blood in PAXgene Blood DNA tubes (QIAGEN # 761115). Alternatively (for smaller or non-human blood samples, an anticoagulant (heparin or EDTA) should be added to blood samples that will be stored. For example, blood samples treated with an anticoagulant can be stored at 2–8°C for a few days or at –20°C or –80°C for a few weeks. Freshly harvested tissue can be immediately frozen and stored at –20°C, –80°C, or in liquid nitrogen. Lysed tissue samples can be stored in a suitable lysis buffer for several months at ambient temperature.
Centrifuge harvested cell cultures, remove the supernatant, and then store the cells at –20°C or –80°C.
Blood samples for RNA: 2.5 mL of whole blood in PAXgene Blood RNA tubes (PreAnalytiX # 762165).
Most biological fluids (e.g., plasma, serum, and urine) and stool samples can be stored at 2–8°C for several hours. Freezing at –20°C or –80°C is recommended for long-term storage. Swabs can be stored dry at room temperature.
It is recommended that RNA samples for Microarray analysis are extracted using the QIAGEN RNAEasy kit with RNAse-free DNase step. Trizol purification is not recommended since residual phenol may inhibit subsequent reactions. The A260/A280 ratio must be between 1.9-2.1 when measured in 10mM Tris pH 7.6. Bioanalyzer profile should not show degradation. A total of 2-5 µg of Eukaryaotic RNA is required.
For standard aCGH experiments, please provide at least 1µg of gDNA (500 ng for 8-plex arrays) for each sample at a concentration of at least 50 ng/mL in either nuclease-free water or 10 mM Tris, pH7.5-8.5.
The following purification kits are recommended (https://www.qiagen.com/us/resources/technologies/ngs/sample-technologies/ )
REAL TIME PCR SAMPLES
Total RNA samples for Real Time PCR must be at least 10 ng/uL with an A260/A280 between 1.7-2.1.
To streamline sample submission, please download our Sample Submission document, edit the appropriate tab(s) in Excel, then attach the updated document and submit. We’ll be in touch with you shortly with your order details.