inhibitor and mimics转染应用说明方法学(国外英文资料).docVIP

inhibitor and mimics转染使用说明方法学(国外英文资料) Mmu-mir-125a-5p research strategy Standard offer Standard offer MicrON? Mmu - miR - 125 - a - 5 p mimic 2 ng 350 RMB 5 ng 600 RMB MicrOFF? Mmu - miR - 125 - a - 5 p inhibitor, 2 ng 300 RMB 5 ng 500 RMB Redye mmu-mir-125a-5p mimic (for example) Contact reebok to synthesize mmu - mir-125a-5p mimic and mmu-mir-125a-5p. The synthetic 2ng mmu-mir-125a-5p mimic USES 1000ul RNase - free water to make 20uM storage liquid, which is packed into 5ul tubes for 30 or -80 freezer (the working concentration of the mimic is 50nM); The day before the transfection, the pancreatic enzyme digests the cells and counts 1X105 c-kit - the cells that make it 90 percent denser. Cells are laid out in half ml of serum, without the normal growing medium of antibiotics. 4. For each hole cells, the use of 50 mu l serum free medium (such as OPTI - MEM) Ⅰ medium dilution 1.25 ul mmu - miR - 125 - a - 5 p mimic liquid storage; 5. For each hole cells, the use of 50 mu l OPTI - MEM Ⅰ serum free medium dilution 1 mu l LIPOFECTAMINE 2000 reagent. Lipofectamine 2000 is diluted with room temperature for 5 minutes (mixed with diluted RNA in 30 minutes). Room temperature is too long to reduce activity. Note: even if Lipofectamine 2000 use the OPTI - MEM Ⅰ diluted, cells can also be use D - MEM cultivation. If d-mem is the diluent of Lipofectamine 2000, the diluted DNA must be mixed within five minutes Mixed diluted RNA (step 4) and diluted Lipofectamine 2000 (step 5). Keep warm for 20 minutes at room temperature. Note: the solution may be cloudy, but it will not affect the transfection. The complex can be stable at room temperature for six hours Directly add the complex into each hole, shake the board, gently blend, add 500ul medium, mix well. Note: if the serum is transfected without serum, the cells can be made using the normal growth medium of the serum. Remove the growth medium from the addition of the complex and replace 0.5 ml without serum medium 8. At 37 ℃ and