DNA Isolation and Banking

WSLH Department: Cytogenetics
WSLH Test Code: 862
Availability: Monday-Friday 7:45 AM - 4:30 PM, Saturday 7:45 AM - 12:00 PM
Turn-around Time: Approximately 2-4 days. (Reports are issued Monday-Friday 7:45 AM - 4:30 PM)
Recommended Uses: Specimens can be used for analysis of genomic DNA sequence by several different methods including but not limited to: PCR based methods, Southern Blotting, restriction digest, and DNA sequencing.
Additional Tests Performed:
Patient Preparations:
Specimen Requirements: **Peripheral blood (including cord blood): 6 ml (preferred) - 2 ml (minimum) in sterile EDTA (purple top) vacuum type tubes.
**Cultured cells: 2 T-25 flasks of cultured cells (80% confluent) from tissues, products of conception (POC), Chorionic Villus Sampling (CVS), or amniocytes.
**Chorionic Villus Sampling: 10mg chorionic villi
**Amniotic fluid: 10-30mL
**Fresh/Frozen tissue: Sample size at least 0.3 cm cubed.
**Skin biopsies: 1-2mm punch biopsy.

Extractions will be attempted on all submitted specimens.
Specimen Handling & Transport: Store and transport specimens at room temperature (may transport with coolant during hot weather, >85 degrees F). DO NOT FREEZE blood samples. The laboratory must receive specimens within 24-48 hours of collection.
Collection Kit/Container: Cytogenetics and Molecular Genetics Collection Kit
Collection Instructions: **Blood: Draw blood using aseptic techniques into a sterile EDTA vacuum type tube(s). Invert tube(s) to mix. If using larger tubes, draw to full volume to avoid over-treatment with anticoagulant.

**Products of conception specimens: Tissue that is pale tan to brown should be avoided if possible as this indicates necrosis. A 0.3-0.5cm cubed section of each tissue type should be collected using aseptic procedures. Place the specimen in a sterile tissue vial containing transport media. If multiple tissues are sent, please place the placenta in one vial and the other tissue(s) in a separate vial to minimize contamination of the tissues.

**Chorionic villus sample: Using aseptic technique, obtain at least 10mg of chorionic villi, taken between 9-38 weeks of gestation. Place villi into a flask with transport media provided by the department.

**Amniotic Fluid: Collect amniotic fluid under sterile, ultrasound guided conditions using a 22-gauge needle inserted through the uterine wall and into the amniotic cavity. Discard the first 1-2ml of amniotic fluid to minimize the possibility of maternal cell contamination. Dispense amniotic fluid into a sterile 15 ml centrifuge tube.

**Skin biopsies: Skin should be cleaned using alcohol, do not use iodine or betadine as these will compromise the cell growth in culture. Do a 1-2mm punch biopsy that goes full depth through the epidermis into the sub-cutaneous fat. Place the specimen in a sterile tissue vial containing transport media.
Unacceptable Conditions: Blood must not be frozen. Plasma and serum are not acceptable. Sample must not be processed with formalin.
Requisition Form: DNA Banking
Required Information: DNA Banking Request Form. Payment due upon request. Laboratory regulations require the following minimum information to be provided on the requisition form for a specimen to be accepted for testing: Patient name or unique identifier; date of collection, patient date of birth and sex, specimen type/site of collection, test request(s). Please be certain that name/identifier on the form matches that on the specimen label.
Results Include: Amount of DNA extracted from specimen available for future testing.
Limitations: **This is NOT intended for Cord Blood Banking or stem cell banking**
**There is a possibility of Maternal Cell Contamination in prenatal specimens**
Additional Tests Recommended:
Additional Comments: Forms that can be downloaded from http://www.slh.wisc.edu/clinical/cytogenetics/ include DNA Banking Request Form, DNA Banking Policy Statement, Request for Release of Specimen, and Transfer of Ownership
Methodology: Extraction of whole genomic DNA and long term storage at -80 degrees. The Maxwell Rapid Sample Concentrator (RSC) Instrument is the platform used for automated purification of nucleic acid from a range of sample types. Using prefilled reagent cartridges, the purification methods use sample lysis and binding to paramagnetic particles as the primary separation principle. DNA is solubilized in a buffered solution containing a DNA stabilizer.
Includes: Isolation of genomic DNA from specimen and long-term storage of DNA.
CPT Code: 81479
Price: For pricing information, please call 608-262-0402.
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