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 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.

 

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