| RNA islotion kits: |
1-1, 1-4 ug, 10-15 ug, 100 ml, 200 ml, 3-4 ug, 5-7 ug, 50 ml, 7-10 ug, A, Add 0, Add at least 1 ml of 75% ethanol per 1 ml of the RNA STAT-60TM used for the initial homogenization, An incubation for 10-15 minutes at 55-60oC may be required to dissolve RNA samples, At the end of the procedure, B, Cells grown in suspension are sediment then lysed in the RNA STAT-60TM (1 ml per 5-10 x 106 cells) by repetitive pipetting, Cellular extraction Cells grown in mono layer are lysed directly in a culture dish by adding the RNA STAT-60TM (1 ml/3, Centrifuge the homogenate at 12, Chlorophom extractions will leave the total negatively charged RNA in the hudrophillic H2O while DNA and proteins are in the hydrophilic, Cs-110, Cs-111, Cs-502 500 ml, Diethylpyrocarbonate (DEPC) treated RNase-free solutions1 should be used for solubilizing of RNA, Dissolve the RNA pellet in water or in 1 mm EDTA, Do not use the Speed-Vac for drying, Following centrifugation, Homogenization RNA STAT-60TM (1 ml per 60-110 mg tissue, Homogenize tissues samples in the RNA STAT-60TM (1 ml/50-100mg tissue) in a Silica +Glass-Teflon homogenizer, Isopropanol extraction and chlorophorm aquouse resuspention is used to yield the RNA in less than 1 hour time withRNA STAT-60TM for total RNA isolation by the RNA STAT-60TM with full intact and protein and DNA free RNA for PCR, It is important not to let the RNA pellet dry completely as it will greatly decrease its solubility, Next, Northern analysis, RNA EXTRACTION Following homogenization, RNA Extraction 1 volume of homogenate + 0, RNA PRECIPITATION Transfer the aqueous phase to a fresh tube and mix with isopropanol, RNA Precipitation 0, RNA STAT-60TM contains phenol and Guanidinium thiocyanate for dissolving with RNA STAT-60TM and a Glass-Teflon or Polytron homogenizer, RNA STAT-60TM is supplied in bottles of 100 ml or 200 ml with red liquid, RNA STAT-60TM islolates one step for RNA from human tissues, RNA isolates more RNA from less specimen biopsy, RNA precipitate (often visible before centrifugation) forms a white pellet at the bottom of the tube, RNA remains exclusively in the aqueous phase whereas DNA and proteins are in the interferes and organic phase, RNA washing Remove supernatant and wash the RNA pellet once with 75% ethanol by vortex and subsequent centrifugation at 7, RNA-STAT60 or RNA STAT-60 from Teltest is used to isolate in less than 1h your total RNA and messenger RNA for PRC, RNASTAT-60, RNase protection assay, RT-PCR and Northern blot, Sample volume should not exceed 10% of the volume of the RNA STAT-60TM used for homogenization, Store samples at room temp for 5-10 minutes and, TISSUE RNA extraction from Tel-test, The final preparation of total RNA is free of DNA and proteins and has a 261/282 ratio, The volume of the aqueous phase is about 60% of the volume of RNA STAT-60TM used for homogenization, Very small samples can be used, Vortex or pass the pellet a few times through a pipette tip, Washing calls before addition of the RNA STAT-60TM should be avoided as this increases the possibility of mRNA degradation, We offer RNA isolation volumes of Cs-112, add 0, and viral origin, animals, b, bacteria, brain, cells, centrifuge at 12, cover the sample tightly, dot blot hybridization, dry the RNA pellet briefly by air-drying or in a vacuum (5-10 min, exosmic mRNA profiling without the use of DNases, fibroblasts, in vitro translation, kidney, molecular cloning, of chloroform, of isopropanol and RNA Wash 75% ethanol, or 0, or 6-10 x 10**6 cells, pH 7, placenta, plants, poly A+ selection, shake vigorously for 15 seconds and let it stay at room temperature for 2-3 minutes, skeletal muscles, spleen, store the homogenate for 5 min at room temp to permit the complete dissociation of nucleoprotein complexes, the homogenate separates into two phases: a lower red phenol chloroform phase and the colorless upper aqueous phase, tissue or exosomic RNA, yeasts, ), ) Cultured cells (ug/10 6 cells): epithelial cells, ) Tissues (ug/mg tissue): liver, ) for 10 min at 4oC, ) for 5 min at 4oC, 000g (max, 000g (max) for 15 minutes at 4oC, 2 ml of chloroform per 1 ml of the RNA STAT-60TM , 2 vol, 5 ml of isopropanol per 1 ml of the RNA STAT-60TM used for homogenization, 5 ug, 5 vol, 5% SDS solution, 500g (max, 5cm petri dish) and passing the cell lysate several times through a pipette, EXPECTED YIELD AND PURITY Expected yield of total RNA: a, RNASTAT60 |