DNA, RNA and protein extraction from fresh frozen tissue or cell pellet
- Solid tissue or cultured cell samples should be processed immediately or else frozen to minimize the activity of endogenous nucleases.
- Slice tissue into pieces no more than 5 mm thick. Cells should be pelleted. Samples must be immediately snap-frozen in liquid nitrogen or in a cryostat.
- If morphologic preservation is not needed, then place the sample in a –80°C freezer indefinitely until DNA or RNA isolation.
DNA/RNA extraction from FFPE and tumor tissue dissection
- Starting material for DNA purification should be freshly cut sections of FFPE tissue. In general, 3 sections (surcharges will be applied to additional slides), each with a thickness of up to 10 μm and a surface area of up to 250 mm2, can be combined in one preparation.
- Tumor tissue can be dissected if the tumor tissue area is marked using the H&E slide as a template.
- Our main technical platform for DNA/RNA extraction from FFPE tissues is the revolutionary Purigen Ionic Purification System that enables automated, column-free, charge-based extraction of nucleic acids with dramatically increased yields and improved purity. Samples are gently lysed and then loaded into the Ionic™ Fluidics Chip. The Ionic system then applies an electric field to the chip and the nucleic acids are isolated in their natural, native form using the company’s proprietary isotachophoresis (ITP) technology.
DNA extraction from saliva collected in Oragene containers
- The recommended volume of saliva is 2 ml. The volume of the Oragene® DNA solution in the kit is 2.0 ml. The total volume of 2 ml of saliva plus the Oragene® DNA solution should be 4.0 ml.
- The full saliva sample should be collected within 30 minutes and the Oragene® DNA kit should be capped immediately. Waiting longer than 30 minutes may decrease the yield and quality of the DNA.
- Mix the Oragene® DNA/saliva sample in the Oragene® DNA vial by inversion.
DNA/RNA extraction from blood-derived samples
- If possible, collect blood in EDTA tube to reduce DNA degradation. However, other anticoagulants such as ACD (citrate) may also be used successfully.
- For best results, the blood should be stored at 4°C for less than 5 days or be frozen at -80°C on the same day.
- For long-term storage of plasma/serum, liquid nitrogen is preferred. We provide service for circulating nucleic acid extraction.
Whole blood processing for lymphocytes and plasma
- Whole blood should be preserved in ACD tubes and EDTA tubes.
- For plasma and whole blood, completely fill the Vacutainer whenever possible to eliminate dilution from the anticoagulant or preservative and immediately mix the blood by gently and thoroughly inverting the tube five to ten times.
DNA quality check by gel electrophoresis
- DNA should be stored in TE to reduce the chance of degradation. Samples can be analyzed with concentrations as low as 5 ng/ul.
DNA quantitation by Picogreen assay
- DNA should be stored in TE or low TE and protected from multiple freeze-thaws as this will cause fragmentation resulting in low Picogreen concentrations.
- Due to the nature of this assay (only double-stranded DNA is detected), Picogreen readings will often result in significantly lower concentrations when compared to spectrophotometric (260/280) readings. The DNA quantitation by Picogreen Assay is necessary for sensitive downstream applications such as arrays and next generation sequencing.
DNA/RNA quality check using TapStation
- We use the Agilent 4200 TapeStation system to check DNA integrity number (DIN) and RNA integrity number (RIN), which are critical for next generation sequencing and other downstream assays.
|BEF Services||2021 Price|
|DNA extraction from blood||$30.00|
|DNA extraction from saliva||$22.00|
|Simultaneous DNA/RNA/protein extraction||$95.00|
|RNA from fresh frozen tissue/FFPE/cells||$50.00|
|DNA from frozen tissue/cells||$28.00|
|DNA from FFPE tissue (up to 3 slides/sample)||$38.00|
|# of additional FFPE slides/Nanodrop readings||$0.50|
|Plasma processing from blood samples||$6.00|
|RNA/DNA QC (RIN/DIN - TapeStation 4200)||$8.00|
|Protein extraction from fresh frozen tissue||$18.00|
|Simultaneous RNA/DNA from tissue/FFPE/cells||$72.00|
|Circulating NA extraction from plasma||$42.00|