Sample Preparation

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.

Microarray Samples

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.

Sequencing Samples

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.

PacBio Samples

Please review the following documents for DNA or RNA (IsoSeq) sample submission and shipping guidelines.

Sample Guidelines\Pacbio Guidelines – DNA v2.0.pdf
Sample Guidelines\Pacbio Guidelines – Iso Seq v2.0.pdf 
Sample Guidelines\Pacbio Guidelines – DNA Shipping Handling v2.0.pdf

BioNano Optical Mapping Samples

The minimum sample amounts required is two Eppendorf tubes with 50 to 100 mg of tissue. MOgene LC will carry out DNA isolation using protocols provided by BioNano Genomics.