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. |
| Contraindications: | |
| 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. |