Protocol for Performing a Skin Biopsy - CENTOGENE

May 2010Centogene GmbHSchillingallee 6818057 Rostock / GermanyTel.: +49 (0)381 203652 0Fax: +49 (0)381 203652 19Mail: info@centogene.deWeb: www.centogene.deHow to perform a skin biopsy for cultivation of fibroblasts for further geneticor biochemical analysis:1. Performing a skin biopsyYou may anesthesize locally if the patient is highly sensitive to pain. For local anesthesia useeither Emla-band-aid 1.5 hours before planned skin biopsy (preferred in children) or afterdisinfection has been performed (directly before skin biopsy) Scandicain 0.5%.The biopsy has to be performed under sterile conditions! The site of biopsy (e.g. axilla,underarm or gluteal) has to be disinfected with ethanol for 10 to 15 minutes. For disinfectionwipe the site of biopsy from the centre in circular movements to the outer site using a sterilepad soaked with 70% ethanol. Take 30 seconds for this procedure and repeat it at minimum5 times using a new ethanol pad each time.Skin biopsy is performed with a sterile skin punch. A pair of tweezers and a scalpel may helpduring biopsy. The punch is held between the thumb and index finger, placed upon the uppersurface the skin and pressed down clockwise in a “screwing” motion at moderate pressure.Caution: do no punch too deep into the subcutaneous tissue, since you do not want toomuch of the subcutaneous fatty tissue. If you perform a deep skin biopsy make sure that theepithelial cover does not remain in the punch.The skin cast punched out will lift itself above the level of the surrounding skin. It should becarefully grasped and raised with a small anatomical set of tweezers, avoiding any crushing.It is then separated away flat with the scissors or scalpel at the base.Transfer the skin biopsy immediately into a sterile 10 ml plastic tube containing 10 ml ofpreferably fresh sterile tissue culture medium (DMEM or RPMI 1640, supplemented with 10%FCS) or otherwise fresh sterile physiologic sodium chloride solution (0.9% NaCl) at roomtemperature (no cooled solutions). Please make sure that the tube is filled up completelyand sealed tightly. Label the tube with the date of biopsy, the name and DOB of the patient.Due to tensioning of the skin, an oval-shaped blemish is produced, which can easily be closedup. Hæmostasis using any instrument is almost never needed. It is easy to dispense withclosing up the wound. Covering up the wound takes place either by compressing or with a smallhydrocolloid tape on the basis of the small tapes which are part of the kit. The resulting scarsare scarcely visible.Managing Director: Christoph Ehlers Deutsche Apotheker- u. Ärztebank Rostock Deutsche Bank FreiburgMedical Director: Prof. Dr. A. Rolfs Code: 300 606 01 Code: 680 700 30Place of jurisdiction: Account: 0006884695 Account: 041357500Amtsgericht Rostock HRB 10296 IBAN: DE56 3006 0601 0006 8846 95 IBAN: DE93 6807 0030 0041 3575 00VAT: DE250191614 BIC/SWIFT: DAAEDEDD BIC/SWIFT: DEUTDE6FXXX

2. Storage of sampleThe sample should be used to start the fibroblast culture at the earliest possible point of timewithin 3 days of the biopsy. Therefore it should not arrive later than 2-3 days in our lab afterthe biopsy has been taken.Store and transport sample at room temperatureMake sure that the sample does not freeze at any time (cover/wrap tube with the skin biopsyusing insulation material).3. Shipment of skin biopsyMake sure that the sample does not freeze during transport (cover/wrap tube with the skinbiopsy using insulation material).Send the skin biopsy immediately at room temperature to the following address:Centogene GmbHInstitute of Molecular DiagnosticsSchillingallee 6818057 RostockTel.: +49 381 2036 52 0FAX : +49 381 2036 5219GermanyManaging Director: Christoph Ehlers Deutsche Apotheker- u. Ärztebank Rostock Deutsche Bank FreiburgMedical Director: Prof. Dr. A. Rolfs Code: 300 606 01 Code: 680 700 30Place of jurisdiction: Account: 0006884695 Account: 041357500Amtsgericht Rostock HRB 10296 IBAN: DE56 3006 0601 0006 8846 95 IBAN: DE93 6807 0030 0041 3575 00VAT: DE250191614 BIC/SWIFT: DAAEDEDD BIC/SWIFT: DEUTDE6FXXX