PCR Sample Preparation
Please use corresponding instructions according to sample types:
1. Sample Treatment
A. RNA Sample: Take 10µg total RNA and provide RNA quality test image. It is required that the concentration is above 500ng/µl, volume is above 20µl and the ratio of OD260/OD280 shall be between 1.8 and 2.0. The lightness of 28s electrophoresis detection is double 18s. Please store at -80°C with dry ice transportation.
B. Tissue Sample: Put over 200mg tissue samples into two frozen pipes and mark. Store at -80°C or in liquid nitrogen with dry ice transportation.
C. Blood Sample: Please provide at least 2mL blood sample, process with 2 hours, add Trizol and mark.
1) Whole Blood: Collect fresh blood and it is suggested to extract RNA by using RNA Isolation Kits for whole blood samples such as QIAamp RNA Blood Mini Kit or separate white blood cells by fresh blood. Place the processed sample into dry ice or store at -80°C refrigerator with dry ice transportation.
2) Plasma: Use sample processed by EDTA anticoagulant, centrifuge for 5 minutes at 4000rpm at 4°C. The upper layer is plasma sample. Add adequate Trizol in the ratio of 1ml Trizol for every 200µl plasma sample, vortex shake and put into dry ice or store at -80°C refrigerator with dry ice transportation.
3) Serum: Collect blood sample by using non-anticoagulant tube and statically isolate the upper layer of serum. Add adequate Trizol in the ratio of 1ml Trizol for every 200µl serum, vortex shake and put into dry ice or store at -80°C refrigerator with dry ice transportation.
D. Cell Samples
a. Adherent Cells
1) After the adherent cell culture or treatment, discard culture media, wash with pre-cooled PBS once;
2) Add Trizol reagent in the ratio of 1mL Trizol reagent for each 10 cm2 culture area and mix gently to ensure all cells are thoroughly digested by Trizol;
3) Repeatedly beat cells by using 1ml pipette tip, transfer Trizol suspension cells into EP tube with RNAase-free and mark well;
4) Store them in dry ice or at -80°C refrigerator with dry ice transportation.
b. Suspension Cells
1) Centrifuge to discard culture medium after the suspension cell culture or drug administration;
2) Quickly wash the collected cells by using pre-cooled PBS buffer solution once and centrifuge to discard PBS buffer solution;
3) Add Trizol reagent in the ratio of 1 mL Trizol for each 5×106 cells, thoroughly suspend cells with vortex oscillation and mark well;
4) Store them in dry ice or at -80°C refrigerator with dry ice transportation.
Note: Clients are required to pay ¥60/each sample for earlier stage processing if providing B, C, D samples.
2. Sample Information Card
1) Please carefully complete the《 “PCR Sample Information Card” 》，and provide sample reference number, quantity, treatment methods, etc.
2) For RNA, please also provide related RNA data and quality test image including spectrophotometry, gel electrophoresis image or Nanodrop detection data.
Note: Information on sample information card shall be consistent with the ones on the contract. Otherwise, all information shall be listed accordingly.
3. Sample Package and Labeling
1) It is suggested to use 1.5ml centrifuge tube to pack samples and seal with sealing film.
2) Make clear sign on each sample tube.
a. It is preferred to mark the product sign on the tube and tube cap using black marker;
b. Write the mark on the paper label and paste on the tube.
Note: Mark on the tube shall be consistent with the ones on the sample information card.
4. Sample Transportation
1) RNA and DNA samples with dry ice transportation.
2) Cell and tissue samples with dry ice transportation.
Note: Dry ice transportation shall not exceed 72 hours.
