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| RNA Sample Collection and Submission Guidelines |
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| Services | RNA Specimen Processing Overview | RNA Extraction FAQ | Sample Collection and Submission Guidelines | Methods and Data Delivery | Scheduling and Shipping |
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Please contact GPCL personnel to clarify procedures before beginning sample collection. Sample Collection In order to maximize the probability of success in your goals we recommend that you follow some basic procedures during sample collection to ensure RNA quality. Tissue Collection Tissue must be protected from degradation and cross contamination throughout the collection process. Gloves should be worn at all times and any instrument used in removing or handling the tissue must be well cleaned and free of even residual material from any other subject or tissue specimen. If the tissue needs to be split it should be rapidly cut with a clean blade. The tissue should be quickly deposited into an appropriate clean container and frozen in liquid nitrogen. If prompt storage in liquid nitrogen is not possible, specimens may be preserved in RNALater (Ambion). Specimens can be minced to <0.5 cm as needed with a clean blade then quickly submerged in 5-10 volumes of RNALater solution. Per RNALater manufacturer's specifications, the RNA is protected from degradation for 24 hours at 37°C, 1 week at 25°C, or 1 month at 4°C. For archival storage the samples should be incubated at 4°C overnight then transferred to -20 or -80°C.The RNALater MUST be removed prior to freezing! This is a particularly desirable method for small biopsies taken in a clinic setting where liquid nitrogen is inaccessible or inconvenient or for shipping specimens overseas where there is risk of delays. Tissue Storage Containers Tissue specimens brought to the Core Laboratories should be put into a cryovial with a screw-top lid. If specimens will not fit in a cryovial, please contact the GPCL for best collection and submission practices. The preferred handling and preservation method for most samples is snap freezing in liquid nitrogen with subsequent transportation on dry ice and long term storage at -80°C. Blood Collection Blood for RNA extraction should be drawn into a PaxGene (Qiagen) or Tempus (Applied Biosystems) tube. It is critical to RNA quality and yield that tubes be thoroughly mixed by inversion at the time of collection, that a full tube of blood be taken and that nothing is placed over the black fill mark on the manufacturer's label. Please be sure the collecting personnel have been thoroughly briefed in collection procedures. Tubes can be refrigerated for up to one week or frozen at -80°C for up to a year before being delivered to the GPCL for processing. For further details see Shipping/Scheduling instructions. Blood Tube Labels All sample tubes must be labeled with the following information in order to be processed: the Study Name, PI Name, Study Reference ID, and the Date Collected. Any other pertinent information regarding the sample must be noted, i.e. infectious status (if known). Sample labels must NOT contain any information revealing the identity of clinical subjects. Please use adherent labels that do not appreciably enlarge the diameter of the tubes so that they can be placed into a centrifuge as needed. See notification section in Shipping/Scheduling for further details. Storage Containers for Mail Delivery Blood samples must be packaged in containers that conform to postal regulations for biohazardous materials and mailed OVERNIGHT to the GPCL Specimen Processing Core Facility for Next day delivery. Frozen tubes must be shipped in dry ice. Cells Cell culture source cells may be provided by the Investigator for RNA isolation. Minimum handling is recommended to avoid changing the expression profile of living cells. Cells grown in suspension should be pelleted, media completely removed and snap frozen. No wash is necessary. Adherent cells should be lysed with TRIzol (Invitrogen) and the lysate submitted for processing. Please contact GPCL personnel for clarification on any of these procedures. Sample Log Sheet A Specimen Processing drop off form must be received with all samples. Studies involving multiple drop offs over a period of months may use a project specific sample log sheet as arranged during the project initiation meeting. Standard drop off forms are available at: http://www.genetics.pitt.edu/forms/ Labels for Containers All sample tubes must be labeled with the following information in order to be processed: the Study Name, PI Name, Study Reference ID, Date collected. Sample labels must NOT contain any information revealing the identity of clinical subjects. All tubes/containers should be labeled with printed labels as laboratory reagents frequently dissolve most marker inks. Drop-off Samples may also be dropped off at one of our sample drop off locations, 3343 Forbes Ave Room 303, and S534A Scaife Hall or 2.15 Hillman Cancer Center Research Pavilion. Frozen tissue samples should be delivered to 3343 Forbes Avenue on dry ice to avoid potential thawing. |