How are samples stored and logged?

The DNA samples are stored at 4 degrees; samples stored on a long term basis are stored at -20 degrees. Currently sample tracking information is stored in Excel files. The data will be transferred to the 'Progeny' database system.
How can I maximize the chance of successful DNA isolations and cell cultures?

The blood should reach the lab 48 hours after being drawn. Blood should be stored and shipped at room temperature and should not be hemolysed. The blood tubes should be filled to capacity as half filled tubes reduce the viability of the cells.
How should I mail out my samples to the lab?
Do you accept blood, saliva and buccal swab samples only?

At the moment these are all that we accept but please contact us to discuss this further.
I am working on a grant funded study and am unsure that I will be able to meet the Core fees.  Can anything be done about this?

Please contact us and we will be sure to get back to you immediately to discuss fees.
What is the procedure for setting up an account with the Core?

Please email hgrc@columbia.edu to create an account.
Which type of tubes should I use for cell lines and DNA Isolations?

For Cell lines use Yellow-top Vacutainer tubes to collect blood.
For DNA Isolations use Yellow-top or Purple-top Vacutainer tubes to collect blood.

Tubes can be ordered from BD Medical Supplies: www.bd.com
Can you recommend a supplier to obtain kits for sampling?

We recommend the following companies:

Polyfoam Packers/Thermosafe for mailers: www.polyfoam.com

Fisher Scientific for tubes, butterfly needles, sharps disposal: www.fishersci.com

Daigger for gauze, band-aids, alcohol preps: www.daigger.com
How much blood is required for toddlers and adults?

For growing children and adults 3 tubes of blood is suggested.
For toddlers 2 tubes is suggested.
What is the procedure for Cell Cultures?

Mononuclear cells are isolated from anti coagulated blood by centrifugation in a Cell Separation Tube. The Cell Separation Tube has a polyester gel and Ficoll Hypaque solution which acts as a separation media. During centrifugation the polyester gel moves to form a barrier separating the mononuclear cells and plasma from the red blood cells. The mononuclear cells are collected by pipetting the cell layer and washing with Balanced Salt Solution. The washed mononuclear cells are suspended in growth medium. Epstein Barr Virus and CyclosporinA are added to the cells to set up the cell culture. The cells are grown in an incubator at 37 degrees Celsius and 5 % carbon dioxide.
How are Cultures established?

Mononuclear cells contain both B and T cells. EBV immortalizes the B cells. The immortalized B cells are called lymphoblastoids. CyclosporinA suppresses the growth of T cells. Immortalization of the cells should become apparent within 96 hours. The cells are fed with 2 mls of growth media in 8-10 days. Cultures are left for another 7 days and then fed by adding 2 mls of growth media. If the culture is doing well, lots of cell clumps will begin to appear and media becomes very acidic. When culture is well established, the cells are transferred to a bigger flask and grown up to a volume of 50mls. At this point the cells are frozen and stored in Liquid nitrogen freezers.
What are the common problems with DNA Isolations and forming cell lines?

The most common problem is the Hemolysis of Blood. The lysed Red Blood Cells interfere in the separation of mononuclear cells. In addition, blood received more than 48 hours post draw is a problem. Half-filled blood tubes reduce the viability of the cells for cell lines.

What is the purpose of filter paper?

Filter papers are used for the purpose of quality control and should be utilized in the field at the time of the blood draw; which we will process for the cost of $2.00. We recommend VWR Labshop www.vwrlabshop.com for the purchase of filter paper supplies.

How do I describe the HGRC in a grant submission?