For more information, please contact:
Steven Madore, PhD, Director, Molecular Biology Laboratory
Susan Jones, Team Leader [Contact for technical questions]
As you pursue your basic or discovery research questions, Coriell’s Molecular Biology Group can help to design the best custom service using our technologies and expertise. Following is some basic information to begin our collaboration.
Following is some basic information to begin our collaboration:
1. What are the best collection methods for blood, cells, and saliva which are intended for Coriell’s Molecular Biology Laboratory?
We recommend collecting blood in ACD tubes and shipping these to Coriell at ambient temperature by overnight express service.
Saliva (2 mls) can be collected using an Oragene•DNA collection device and product guidelines; item number OG-300, DNA Genotek. Samples can be shipped to Coriell at ambient temperature.
For RNA, blood should be collected in PAXgene tubes, mixed well, and shipped on dry ice by overnight courier. Alternatively, for RNA isolation, “buffy coats” of PBMCs can be prepared and then shipped as frozen cell pellets on dry ice by overnight courier.
Expected Yields of Nucleic Acid
| Sample type | Cell number/Sample volume | Yield in µg of DNA | Yield in µg of total RNA | Preferred Shipping Method |
| Whole blood, ACD tubes | 10 mls | 30 – 60 µgms | Not applicable | Ambient Temperature Overnight |
| PMBCs | 5 – 10 x 106 cells | 30 – 60 µgms | 5 - 20 µgms | Frozen cell pellet on dry ice |
| Whole blood, PAXgene tube | 2.5 mls blood in preservative | Not applicable | >3 µgms | Overnight |
| Cultured cells | 1 x 107 | 60 µgms | 20 – 100 µgms | Frozen cell pellet on dry ice Overnight |
| Saliva in Oragene•DNA | 2 mls saliva + 2 mls reagent | 90- 100 µgms | Not applicable | Ambient Temperature Overnight |
2. What is the best way to send cells for the purpose of Nucleic Acid extraction at Coriell?
Cultured cells grown in suspension should be collected by centrifugation at 1300 x g, washed once with sterile, ice cold PBS, and shipped as a frozen cell pellet overnight on dry ice.
Cells grown as monolayers should be rinsed once in ice cold PBS and then collected by scraping using a rubber policeman. After spinning at 1300 x g and removing the supernatant, the cells should be shipped to Coriell as a frozen cell pellet by overnight courier.
3. How is DNA isolated and what is the quality control program for that process?
DNA is extracted from whole blood or cells pellets using a modified “salting out” procedure on the AUTOPURE LS instrument. RNase is included during the extraction procedure and the DNA is resuspended in 10 mM TRIS, pH 8.0, 1 mM EDTA. DNA is quantified by spectrophotometric absorbance at A260.
Quality is assessed by the A260/A280 ratio [>1.7 and < 2.1] and average size is estimated by agarose gel electrophoresis.
4. How is RNA isolated and what are the quality control steps?
Blood total RNA is prepared from PAXgene tubes using the PAXgene Blood RNA Kit. The protocol can be modified to include miRNAs, as well. RNA yields from 2.5 ml healthy human whole blood are ≥3 μg for ≥95% of the samples processed with A260/A280 values between 1.8 and 2.2.
RNA is prepared from cell pellets using the QIAgen RNeasy Kit.
All RNA is subjected to analysis on the Agilent 2100 Bioanalyzer and must have a RIN value greater than 7.5. RNA is quantified by A260 on a Nanodrop spectrophotometer.
Support research at Coriell. Let’s find answers together.
