| RNA islotion kits: |
# T-4025), # T1-3200) this treatment step is unnecessary, (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction, (1989) Molecular Cloning, (1989) The RNAzolTM B method, 0, 1, 1, 1, 1-1, 10-15 ug, 156-159, 2, 2, 2, 2, 2, 2, 3, 3, 3, 3, 3, 3-4 ug, 4, 4, 4, 4, 5, 5, 5, 5, 5-7 ug, 50 ml, 6, 7, 7-10 ug, APPLICATION The total RNA/DNA isolated by RNA-DNA STAT-60 is used for Northern and Southern analysis, Add 0, Add 0, Add 0, Add 0, Add 800 ul of DNA STAT-60 reagent per 1 ml of RNAzol, Add at least 1 ml of 75% ethanol per 0, Add at least 1 ml of 75% ethanol per 1 ml of the DNA STAT-60 used for the initial homogenization, Add at least 1 ml of 75% ethanol per 1 ml of the RNA STAT-60TM used for the initial homogenization, Additional references available upon request, An additional precipitation may be necessary to use RNA isolated by the RNA STAT-60TM in enzymatic assays, An incubation for 10-15 minutes at 55-60 oC may be required to dissolve RNA samples, An incubation for 10-15 minutes at 55-60oC is required to completely dissolve RNA samples, An incubation for 10-15 minutes at 55-60oC may be required to dissolve DNA samples, Anal, At the end of the procedure, Avoid breathing vapor, Avoid breathing vapor, Avoid breathing vapor, B, B, BIOLOGICAL FLUIDS, Biochem, Biotechniques 8:148-149, Boehringer Mannheim) to the aqueous phase is necessary for a 100 recovery of the RNA precipitate, CAN BE FATAL, CAN BE FATAL, CSF, Can be fatal, Cells: Cells grown in mono layer are lysed directly in a culture dish by adding the RNA STAT-60 TM (1 ml per 5-10 x 106 cells) by repetitive pipetting, Centrifugate the homogenate at 12, Centrifuge the homogenate at 2000 g (min, Chomczynski, Chomczynski P, Chomzynski P, Citation of the method: total RNA was isolated by the single-step method (2) using RNA STAT-50TM LS, Cold Spring, Cold Spring Harbor Laboratory, Cs-113, DNA PRECIPITATION Transfer the aqueous phase to a fresh tube and mix with isopropanol, DNA WASH Remove supernatant and wash the DNA pellet once 75% ethanol by vortexing and subsequent centrifugation at 7, DNA remains in the aqueous phase whereas RNA and proteins are in the inter phase and organic phase, Diethylpyrocarbonate (DEPC) treated RNase-free solutions should be used for solubilization of RNA, Dissolve the DNA pellet in water or in 1 mm EDTA, Dissolve the RNA pellet in FORMazolTM (Cat, Dissolve the RNA pellet in water or in 1 mM EDTA, Do not get on skin or clothing, Do not get on skin or clothing, Do not get on skin or clothing, Do not use the Speed-Vac for drying, Do not use the SpeedVac for drying, EXPECTED YIELD AND PURITY Expected yield of total RNA:A, Excellent recovery of RNA/DNA permits the use of this product for isolation of RNA/DNA from very small biological samples (biopsies etc, Following centrifugation, Following centrifugation, Following centrifugation the homogenate separates into two phases: a lower organic phase and the upper aqueous phase, Following homogenization (before addition of chloroform) samples can be stored at -70oC for at least 2 weeks, Following homogenization (before addition of chloroform) samples can be stored at -70oC for at least two weeks, Following solubilization, For isolation of RNA from a small amount of cells or tissue (1-10 mg): homogenize samples in 0, HOMOGENIZATION A, Hand and dust may be the major source of the RNase contamination, Hands and dust may be the major source of the RNase contamination, Harbor, Homogenization 0, Homogenization RNA STAT-60 (1 ml per 50-100 mg tissue, Homogenize 0, If the expected yield of RNA is <, If the sample vol, In case of contact: Immediately flush eyes or skin with a large amount of water for at least 15 minutes and seek immediate medical attention, Isolation of RNA from small samples (<, Isopropanol (ACS grade), It is important not to let the RNA pellet dry completely as it will greatly decrease its solubility, It is important not to let the RNA pellet dry completely as this will greatly decrease its solubility, Lyse (or homogenize) samples in 0, Mix 0, Multi-sample processing is a great advantage of using these simple single reagents, N, NOTE: Should you use our RNase free purified water (Cat, NOTES AND COMMENTS 1, NOTES AND COMMENTS 1, Next, Next add 0, P, PCR without DNAses added, PROTOCOL FOR DNA EXTRACTION FROM SAMPLE USED FOR RNA EXTRACTION DNA REVERSE EXTRACTION Remove aqueous layer containing RNA, PROTOCOL RNA/mRNA isolation by the RNA-DNA STAT-60 method includes the following steps: 1, REAGENTS REQUIRED BUT NOT SUPPLIED Chloroform (ACS grade), REFERENCES 1, REFERENCES 1, RNA EXTRACTION Following homogenization, RNA Extraction 1 vol, RNA Extraction homogenate +0, RNA PRECIPITATION Transfer the aqueous phase to a fresh tube and mix with isopropanol, RNA Precipitation 0, RNA Precipitation aqueous phase +0, RNA STAT-50 TM LS works on blood, RNA STAT-50TM LS does not contains chloroform, RNA WASH Remove supernatant and wash the RNA pellet once with 75% ethanol by vortexing and subsequent centrifugation at 7, RNA Wash 1 ml 75% ethanol, RNA Wash 75% ethanol, RNA precipitate (often invisible before centrifugation) forms a white pellet at the bottom of the tube, 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 inter phase and organic phase, RNAzol B or RNA STAT-60 used for the initial homogenization, RNase protection assay, RNase protection assay, Read also the warning note on the bottle, Read also the warning note on the bottle, Read warning note on bottle, SPECIAL HANDLING PRECAUTIONS The DNA STAT-60TM contains an irritant (guanidinium salts), SPECIAL HANDLING PRECAUTIONS The RNA STAT-50TM LS contains poison (phenol) and irritant (guanidinium thiocyanate), SPECIAL HANDLING PRECAUTIONS The RNA STAT-60TM contains poison (phenol) and irritant (guanidinium thiocyante), Sambrook J, Sample volume should of the RNA STAT-60 used for homogenization, Single Step Method of RNA Isolation by Acid Guanidinium Thiocyanate-Phenol-Chloroform Extraction, Steps: 1, Store samples at room temperature for 5-10 min and centrifuge at 12, Store samples at room temperature for 5-10 minutes and centrifuge at 12, TISSUE SUSPENSIONS, The DNA precipitate forms a small clear to white pellet at the bottom of the tube, The PCR and RNase protection assays do not require this traditional precipitation step, The RNA STAT-50TM LS cotains phenol and Guanidinium, The SDS solution used for RNA solubilization should be made free of RNase by diethyl pyrocarbonate (DEPC) treatment, The final preparation of total RNA is free of DNA and proteins and has a 260/280 ration >, The recovery of undegraded RNA and DNA is 30-150% greater than with any other method of RNA/DNA isolation, The total RNA is extracted with isopropanol, The use of sterile, The volume of the aqueous phase is about 60% of the volume of RNA STAT-60TM used for homogenization, The volume of the aqueous phase is about 60% of the volume of homogenate, The volume ration of RNA STAT-50TM LS to sample should always be 3 to 1, These are the most effective methods for isolation of RNA and DNA isolation from the same sample, Tissues: Homogenized tissue samples in the RNA STAT-60 (1 ml/ 50-100 mg tissue) in a glass-Teflon or Polytron homogenizer, Total RNA in H2O while DNA and proteins are extracted into an organic phase and interphase, Total RNA isolated by RNA STAT-50TM LS is pure without DNA contamination for Northern Blot, Total and exosomic RNA 1 step with RNA STAT50TM LS for total RNA purification from biopsies, Unless stated otherwise the procedure is carried out at room temperature, Use at least 0, Use gloves and keep tubes closed, Use gloves and keep tubes closed throughout the procedure, Vortex or pass the pellet a few times through a pipette tip, Vortex or pass the pellet a few times through a pipette tip, Vortex or pass the pellet a few times through a pipette tip, Washing cells before addition of the RNA STAT-60TM should be avoided as this increases the possibility of mRNA degradation, When working with DNA STAT-60 use gloves and eye protection (shield, When working with RNA STAT-50TM LS use gloves and eye protection (shield, When working with the RNA STAT-60TM use gloves and eye protection (shield, add 0INTRODUCTION The entire procedure for RNA/DNA isolation using RNA-DNA STAT-60 can be completed in less than 90 minutes, adjust the volume with water, amniotic fluid, an addition of 5 ug of carrier tRNA or glycogen (molecular biology grade, and Ethanol (ACS grade), and Maniatis T, and Saachi, and Sacchi N, and polymerase chain reaction from human, animal, animals, brain, centrifuge the homogenate at 12, cover the samples tightly, disposable polypropylene tubes is recommended throughout the procedure, dot blot hybridization, dot-blot hybridization, dry briefly the RNA pellet by air-drying or under vacuum (5-10 min), dry the DNA pellet briefly by air drying or in a vacuum (5-10 min, dry the RNA pellet briefly by air-drying or in a vacuum (5-10 min, fibrolasts, human samples, in vitro translation, in vitro translation, is <, isopropanol and ethanol, kidney, mRNA profiling, molecular cloning, molecular cloning, of chloroform 3, of homogenate +0, of isopropanol, or 0, or 5-10 x 106 cells), pH 7, pH 7, phenol-chloroform phase and the colorless upper aqueous phase whereas DNA and proteins are in the inter phase and organic phase, placenta 1-4 ug, plant without any additional RNase/DNase treatment, plants, poly A+ selection, poly A+ selection, precipitate RNA in the presence of 0, safety goggles), safety goggles), safety goggles), serum, shake vigorously for 15 seconds and let it stay at room temperature for 2-3 minutes, shake vigorously for 15 seconds and let it stay at room temperature for 2-3 minutes, shake vigorously for 15 seconds and let them stay at room temperature for 2-3 minutes, skeletal muscles, spleen, store the homogenate for 5 minutes at room temperature to permit the complete dissociation of nucleoprotein complexes, store the homogenate for 5 minutes at room temperature to permit the complete dissociation of nucleoprotein complexes, the homogenate separates into two phases: a lower red, the homogenate separates into two phases: a lower red phenol chloroform phase and the colorless upper aqueous phase, transfer the homogenate to the eppendorf tube and follow the isolation protocol with the exception of the RNA precipitation which should be carried out for 30 minutes at 4 oC, washed with ethanol and water dissolved, water or in 0, yeasts and bacteria, ), ), ) - 12, ) Cultured cells (ug/106 cells): epithelial cells, ) REAGENTS SUPPLIED RNA STAT-60 and DNA STAT-60Preparation: Ready to use, ) Tissues (ug/mg tissue): liver, ) for 10 minutes at 4oC, ) for 10 minutes at 4oC, ) for 10 minutes at 4oC, ) for 15 minutes at 4oC, ) for 15 minutes at 4oC, ) for 15 minutes at 4oC, ) for 5 minutes at 4oC, ) for 5 minutes at 4oC, ) for 5 minutes at 4oC, 000 g (max, 000 g (max, 000 g (max, 000 g (max, 000 g (max, 000 g (max, 1 HOMOGENIZATION A, 1 ug, 106 of cells), 2 M NaCl with two volumes of ethanol for 15 minutes at 4oC, 2 RNA EXTRACTION Following homogenization, 2 ml chloroform, 2 ml of chloroform per 0, 2 ml of chloroform per 1 ml of DNA STAT-60, 2 ml of chloroform per 1 ml of the RNA STAT-60TM cover the sample tightly, 2 vol, 25 ml, 25 ml of sample and lyse cells (or cellular debris) suspended in the sample by passing the mixture several times through a pipette, 25 ml sample, 25 ml sample with 0, 3 RNA PRECIPITATION Transfer the aqueous phase to a fresh tube and mix with isopropanol, 4, 4 RNA Wash Remove supernatant and wash the RNA pellet once with 75% ethanol and subsequent centrifugation at 7, 5 ml isopropanol, 5 ml of isopropanol per 0, 5 ml of isopropanol per 1 ml of the DNA STAT-60 used for homogenization store samples at room temperature for 5-10 minutes and centrifuge at 12, 5 ml of isopropanol per 1 ml of the RNA STAT-60TM used for homogenization, 5 ug, 5 vol, 5% SDS solution, 5% SDS solution, 500 g (max, 500 g (max, 500 g (max, 75 ml RNA STAT-50TM + 0, 75 ml of RNA STAT-50 TM LS for homogenization, 75 ml of RNA STAT-50TM LS, 75 ml of RNA STAT-50TM LS in a glass-Teflon or Polytron homogenizer, 75 ml of RNA STAT-50TM LS in the Eppendorf tube and follow the isolation protocol with the exception of the RNA precipitation which should be carried out for 30 minutes at 4oC, 75 ml of RNA STAT-50TM LS per 5-10 x 106 cells, 75 ml of RNA STAT-50TM LS used for the initial homogenization, 75 ml of RNA STAT-50TM LS with 0, 8, 8, 8, 162, 1987, 8 ml of the RNA STAT-60TM, At the end of the procedure, At the end of the procedure, B, F, Fritsch E, RNA Stat 50 LS and RNA STAT 50 LS, Stability: Refer to expiration date stamped on label, Storage: Refrigerate at 2-8oC, The final preparation of total RNA is free of DNA and proteins and has a 260/280 ratio >, Y, exosomic microRNAs in 60 minutes |