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Matches in Nanopublications for { ?s <http://purl.org/dc/elements/1.1/description> ?o ?g. }

• step description "Pellet down bacilli by centrifugation at 2,000 x g for 10 min, room temperature and carefully discard supernatant." assertion.
• step description "Bring down temperature of suspension to ~30 °C and add 20 µl of 100 mg/ml lysozyme solution (final concentration, 2 mg/ml) to it. Incubate suspension at 37 °C for 6 h to overnight in a water bath. Note: In case of fast-growing mycobacteria such as Mycobacterium smegmatis (M. smegmatis), go directly to step 7. However, if DNA is to be isolated from slow-growing species, follow steps 4-6 which involve lysis of cells by a physical method. Because of enhanced disruption of cells, this would significantly enhance yield." assertion.
• step description "Harvest mycobacteria by centrifuging 20-40 ml log phase culture (O.D.600 ≈ 0.8) at 2,000 x g for 10 min, room temperature. Carefully discard culture medium and add 10 ml TE (at room temperature) buffer to pellet. Mix gently with a pipette. Note: If isolating DNA from pathogenic mycobacteria such as M. tuberculosis, step 1 and 2 must be performed in a BSL-3 facility." assertion.
• step description "Aspirate liquid without disturbing pellet. Keep tube open to air dry pellet. Do not over dry pellet, as over dried DNA will be difficult to dissolve." assertion.
• step description "Centrifuge tube at 12,000 x g for 10 min, 4 °C. Discard liquid carefully." assertion.
• step description "Repeat steps 6 to 8 for achieving a higher purity of DNA." assertion.
• step description "Add 1 ml of 10% SDS solution (final concentration, ~1%) and 20 µl of 10 mg/ml Proteinase K solution (final concentration 20 µg/ml). Incubate for 2 to 4 h at 60 °C in a hot water bath." assertion.
• step description "The cells in interphase need to be aspirated without delay. If the tubes are kept standing for more than 10 min, PBMCs from the interphase will get disturbed and start settling down." assertion.
• step description "Centrifuge the tubes (without any delay) for 30 min at 100 x g in 4 °C in a swing-out bucket. Fixed angle rotors also can be used but would require more caution when separating cells in interphase." assertion.
• step description "Take 4 ml of Ficoll Histopaque in a 15 ml centrifuge tube." assertion.
• step description "Cells can be treated with different antigens for different period of times and the supernatants can be analysed for cytokine levels. The cells can be analysed for phenotypic change, apoptosis or proliferation. Note: PBMCs are primary cells and cannot be cultured for more than one passage under normal conditions. Lymphocytes of PBMCs can be made to proliferate in vitro by mitogens e.g., Phytohaemagglutin or Concanavin-A etc over a period to 72-96 h. Monocytes generally are end cells and do not proliferate. In absence of mitogens the proliferation of PBMCs will be negligible." assertion.
• step description "Discard medium and re-suspend the cell pellet in 1 ml of sterile Dulbecco's modified eagle medium." assertion.
• step description "Isolate human PBMCs by gradient centrifugation using Ficoll-Histophaque." assertion.
• step description "In this step, incubate samples at 65℃ for 10 min for lysing cells completely." assertion.
• step description "Ground samples (leaf, shoot, root, and recalcitrant samples, approximately 0.5-1 g) using 1 ml of the extraction buffer with or without liquid nitrogen in mortar and pestle that are sterilized.CRITICAL STEP The procedure are carried out at room temperature except the centrifugation steps (at 4℃) as well as the time of precipitating of the nucleic acid using the isopropanol (on ice).CRITICAL STEP Severely disrupt the tissues to create the glaze mode of samples." assertion.
• step description "After centrifuging at 5400 g at 4˚C for 5 min, remove the supernatant and then air-dry the pellet." assertion.
• step description "Centrifuge tubes at 13700 g at 4℃ for 10 min. The white pellet will be visible on the bottom of the tubes.CRITICAL STEP Do not disturb bottom phases of the solution when you pipet the upper phase." assertion.
• step description "Transfer the upper aqueous phase to a new tube (size= 1.5 ml)." assertion.
• step description "Transfer the resulting solution to a sterile centrifuge tube (size=2 ml), and then mix sample by briefly vortexing until the sample is thoroughly resuspended." assertion.
• step description "The IBV-infected cells were incubated at 37 °C in 5% CO2." assertion.
• step description "The RNAs were stored at -80 °C for further use. " assertion.
• step description "The RNA pellets are air-dried and dissolved in 100 µl RNase-free H2O by incubating at 65 °C for 15 min." assertion.
• step description "RNA was precipitated by centrifugation at 12,000 x g for 10 min at 4 °C." assertion.
• step description "The upper aqueous phase was transfer into a new tube and mixed with 1:1 (volume/volume) of 100% isopropanol, and then incubated for 10 min at room temperature." assertion.
• step description "In this step, incubate samples at 65℃ for 10 min for lysing cells completely." assertion.
• step description "Ground samples (leaf, shoot, root, and recalcitrant samples, approximately 0.5-1 g) using 1 ml of the extraction buffer with or without liquid nitrogen in mortar and pestle that are sterilized.CRITICAL STEP The procedure are carried out at room temperature except the centrifugation steps (at 4℃) as well as the time of precipitating of the nucleic acid using the isopropanol (on ice).CRITICAL STEP Severely disrupt the tissues to create the glaze mode of samples" assertion.
• step description "After incubating at room temperature for a few minutes, keep the solubilized nucleic acid in −20˚C for a short time storage or in −80˚C for a longtime storage. Figure 1 shows a schematic model of all the DNA and RNA isolation steps of this procedure." assertion.
• step description "Dissolve the pellet in 20-30 µl of RNase free water (commercial) or autoclaved water." assertion.
• step description "Centrifuge tubes at 13700 g at 4℃ for 10 min. The white pellet will be visible on the bottom of the tubes.CRITICAL STEP Do not disturb bottom phases of the solution when you pipet the upper phase" assertion.
• step description "Transfer the upper aqueous phase to a new tube (size= 1.5 ml)" assertion.
• step description "Grind mosquitoes in 1.5 ml eppendorf tube with micropistle in 50-100 µl 1X STE buffer ( 50mM Nacl,50mM Tris- HCL,100mM EDTA, Ph 8.0) along with 100mM sucrose. Add 1X STE buffer to a total volume of 300-500 µl for a single mosquito and 1 ml for mosquito pool like 4,6,8,10 numbers. Then add 1% SDS ,1% Triton -X , 10 µl/ ml Rnase A (20mg/ml), 20 µl/ ml Proteinase K (20mg/ml) and mix it." assertion.
• step description "Wash preserved mosquitoes in sterile distilled water or phosphate buffer saline(PBS) to remove excess alcohol. Fresh mosquitoes can be ground directly." assertion.
• step description "Keep the pellet at 37 º celcius for 10 minutes" assertion.
• step description "Centrifuge at 12,000g for 30 minutes at 4º celcius and then remove the supernatant" assertion.
• step description "Repeat the above step(5) , then add chloroform:isoamyl alcohol(24:1) and centrifuge at 12,000g for 10 minutes at 4º celcius " assertion.
• step description "Centrifuge at 12,000g for 10 minutes at 4º celcius .Transfer the supernatant to a fresh tube" assertion.
• step description "After incubation, the tubes were centrifuged at 5000 × g for 10 min at 4°C and the supernatant was gently removed. The pellet is washed two times with 1ml of 70% ethanol and the DNA is pellet by 5000 × g at 4°C for only 5 min. The supernatant is discarded and the pellet is air-dried (10 min). The pellet are allowed to re-suspend in 50 μL of TE (10 mM Tris. HCl pH 8.0; 1 mM EDTA pH 8.0)." assertion.
• step description "After RNAse A treatment, add one volume of chloroform: isoamyl alcohol again and mix by inversions for 5 min. The tube is centrifuged for 10 min at 5000 × g at 4°C. Transfer the clear supernatant into new 2ml tube and add half volume of 5 M NaCl to the sample and mix gently by inversions" assertion.
• step description "Transfer 1ml of supernatant to each 2ml Eppendorf tubes already containing 1 ml of chloroform: isoamyl alcohol (24:1). Mix supernatant and chloroform: isoamyl alcohol by gentle inversions for 10 min and subsequently place the tube on ice for 10 min." assertion.
• step description "If in step one the sample is not grinded with mortar and pestle then vortex the falcon tube for 5 min otherwise proceed to step 3" assertion.
• step description "Dissolve pellet in molecular biology grade water and analyze DNA by agarose gel electrophoresis and spectrophotometer for its purity and integrity" assertion.
• step description "Snap freeze pellet in liquid N2 and immediately transfer to a mortar. Using a pestle, crush bacteria while repeatedly adding liquid N2 (~20 ml) to it." assertion.
• step description "Bring down temperature of suspension to ~30 °C and add 20 µl of 100 mg/ml lysozyme solution (final concentration, 2 mg/ml) to it. Incubate suspension at 37 °C for 6 h to overnight in a water bath. Note: In case of fast-growing mycobacteria such as Mycobacterium smegmatis (M. smegmatis), go directly to step 7. However, if DNA is to be isolated from slow-growing species, follow steps 4-6 which involve lysis of cells by a physical method. Because of enhanced disruption of cells, this would significantly enhance yield." assertion.
• step description "Incubate mycobacterial suspension at 80 °C for 1 h in a water bath. This step will facilitate loosening of the mycobacterial cell wall and will also inactivate pathogenic mycobacteria." assertion.
• step description "Aspirate liquid without disturbing pellet. Keep tube open to air dry pellet. Do not over dry pellet, as over dried DNA will be difficult to dissolve" assertion.
• step description "Centrifuge tube at 12,000 x g for 10 min, 4 °C. Discard liquid carefully" assertion.
• step description "Add 100 µl 3 M sodium acetate solution per ml of aqueous phase obtained in step 10. Mix gently by inverting tube" assertion.
• step description "Centrifuge suspension at 12,000 x g for 10 min, 4 °C. Carefully transfer upper aqueous phase to a new 50 ml tube" assertion.
• step description "Wash preserved mosquitoes in sterile distilled water or phosphate buffer saline(PBS) to remove excess alcohol. Fresh mosquitoes can be ground directly." assertion.
• step description "Wash the pellet with 70% ethanol ." assertion.
• step description "Transfer the very clear supernatant to a fresh tube , add two fold volume cold isopropanol and keep it for 1 hour at -20º celcius ." assertion.
• step description "Add equal volume of phenol:chloroform(1:1) ,shake the tube well for 5 minutes and centrifuge at 12,000g for 10 minutes at 4º celcius ." assertion.
• step description "Centrifuge at 12,000g for 10 minutes at 4º celcius .Transfer the supernatant to a fresh tube." assertion.
• step description "After incubation, the tubes were centrifuged at 5000 × g for 10 min at 4°C and the supernatant was gently removed. The pellet is washed two times with 1ml of 70% ethanol and the DNA is pellet by 5000 × g at 4°C for only 5 min. The supernatant is discarded and the pellet is air-dried (10 min). The pellet are allowed to re-suspend in 50 μL of TE (10 mM Tris. HCl pH 8.0; 1 mM EDTA pH 8.0)." assertion.
• step description "After RNAse A treatment, add one volume of chloroform: isoamyl alcohol again and mix by inversions for 5 min. The tube is centrifuged for 10 min at 5000 × g at 4°C. Transfer the clear supernatant into new 2ml tube and add half volume of 5 M NaCl to the sample and mix gently by inversions." assertion.
• step description "Transfer 1ml of supernatant to each 2ml Eppendorf tubes already containing 1 ml of chloroform: isoamyl alcohol (24:1). Mix supernatant and chloroform: isoamyl alcohol by gentle inversions for 10 min and subsequently place the tube on ice for 10 min." assertion.
• step description "If in step one the sample is not grinded with mortar and pestle then vortex the falcon tube for 5 min otherwise proceed to step 3." assertion.
• step description "Take one gram of frozen finely powdered leaf tissues in a new 50 mL Falcon tube and mixed with the pre-heated extraction buffer (10 ml) for one sample. Breifly, vertex for 30sec to ensure leaf material is fully mixed with buffer. Fine grind is a key to obtaining high DNA quantity with lesser artifacts of resin." assertion.
• step description "Cells were seeded in 100-mm-diameter dishes and infected with either 2 PFU of live IBV per cell or the same amount of UV-inactivated IBV (UV-IBV) at 37 °C. Excess virus in the medium was removed by replacing with fresh medium at 1 h post-infection." assertion.
• step description "RNA concentration and purity were determined by NanoDrop." assertion.
• step description "The RNA pellets are air-dried and dissolved in 100 µl RNase-free H2O by incubating at 65 °C for 15 min." assertion.
• step description "Cell lysates were transfer into eppendorf tubes and one-fifth (volume/volume) of chloroform was added to each tube" assertion.
• step description "Cells were seeded in 100-mm-diameter dishes and infected with either 2 PFU of live IBV per cell or the same amount of UV-inactivated IBV (UV-IBV) at 37 °C. Excess virus in the medium was removed by replacing with fresh medium at 1 h post-infection." assertion.
• step description "Transfer the very clear supernatant to a fresh tube , add two fold volume cold isopropanol and keep it for 1 hour at -20º celcius" assertion.
• step description "Wash the pellet with 70% ethanol" assertion.
• step description "Keep the pellet at 37 º celcius for 10 minutes" assertion.
• step description "Grind mosquitoes in 1.5 ml eppendorf tube with micropistle in 50-100 µl 1X STE buffer ( 50mM Nacl,50mM Tris- HCL,100mM EDTA, Ph 8.0) along with 100mM sucrose. Add 1X STE buffer to a total volume of 300-500 µl for a single mosquito and 1 ml for mosquito pool like 4,6,8,10 numbers. Then add 1% SDS ,1% Triton -X , 10 µl/ ml Rnase A (20mg/ml), 20 µl/ ml Proteinase K (20mg/ml) and mix it." assertion.
• step description "Wash preserved mosquitoes in sterile distilled water or phosphate buffer saline(PBS) to remove excess alcohol. Fresh mosquitoes can be ground directly." assertion.
• step description "Wash the pellet with 70% ethanol ." assertion.
• step description "Keep the pellet at 37 º celcius for 10 minutes." assertion.
• step description "Dissolve the dry pellet in nuclease free water or TE buffer (Ph 8.0). Store DNA at 4 º celcius or -20 º celcius ." assertion.
• step description "After RNAse A treatment, add one volume of chloroform: isoamyl alcohol again and mix by inversions for 5 min. The tube is centrifuged for 10 min at 5000 × g at 4°C. Transfer the clear supernatant into new 2ml tube and add half volume of 5 M NaCl to the sample and mix gently by inversions" assertion.
• step description "After incubation, the tubes were centrifuged at 5000 × g for 10 min at 4°C and the supernatant was gently removed. The pellet is washed two times with 1ml of 70% ethanol and the DNA is pellet by 5000 × g at 4°C for only 5 min. The supernatant is discarded and the pellet is air-dried (10 min). The pellet are allowed to re-suspend in 50 μL of TE (10 mM Tris. HCl pH 8.0; 1 mM EDTA pH 8.0)." assertion.
• step description "The IBV-infected cells were incubated at 37 °C in 5% CO2" assertion.
• step description "Cell lysates were transfer into eppendorf tubes and one-fifth (volume/volume) of chloroform was added to each tube." assertion.
• step description "The upper aqueous phase was transfer into a new tube and mixed with 1:1 (volume/volume) of 100% isopropanol, and then incubated for 10 min at room temperature." assertion.
• step description "RNA was precipitated by centrifugation at 12,000 x g for 10 min at 4 °C." assertion.
• step description "The RNA pellets are air-dried and dissolved in 100 µl RNase-free H2O by incubating at 65 °C for 15 min." assertion.
• step description "RNA pellet was washed with 1 ml 70% RNase-free ethanol once and spin down by 7,500 x g for 5 min." assertion.
• step description "@FairStep(fw) def add(dog, cat): """ Add two integers together (dog and cat). """ result = dog + cat return result " assertion.
• step description "@FairStep(fw) def mult(walrus, bird): """ Multiply two integers together (walrus and bird). """ result = walrus * bird return result " assertion.
• step description "@FairStep(fw) def add(dog, cat): """ Add two integers together (dog and cat). """ result = dog + cat return result " assertion.
• add description "@FairStep(fw) def add(int1, int2): """ Add two integers together (int1 and int2). """ result = int1 + int2 return result " assertion.
• mult description "@FairStep(fw) def mult(int1, int2): """ Multiply two integers together (int1 and int2). """ result = int1 * int2 return result " assertion.
• mult description "@FairStep(fw) def mult(int1, int2): """ Multiply two integers together (int1 and int2). """ result = int1 * int2 return result " assertion.
• add description "@FairStep(fw) def add(int1, int2): """ Add two integers together (int1 and int2). """ result = int1 + int2 return result " assertion.
• mult description "@FairStep(fw) def mult(int1, int2): """ Multiply two integers together (int1 and int2). """ result = int1 * int2 return result " assertion.
• RA6TLVO9Rz8G5hLf139JpfjRl1lgLz54-3FUdXtnUaNUg description "@FairStep(fw) def mult(walrus, bird): """ Multiply two integers together (int1 and int2). """ result = walrus * bird return result " assertion.
• RAuUwfI-EVDpBG-roDmKNTGoe0t_Y7p90d362XicpK4xc description "@FairStep(fw) def add(dog, cat): """ Add two integers together (int1 and int2). """ result = dog + cat return result " assertion.
• step description "@FairStep(fw) def add(dog, cat): """ Add two integers together (int1 and int2). """ result = dog + cat return result " assertion.
• step description "@FairStep(fw) def mult(walrus, bird): """ Multiply two integers together (int1 and int2). """ result = walrus * bird return result " assertion.
• step description "@FairStep(fw) def add(dog, cat): """ Add two integers together (dog and cat). """ result = dog + cat return result " assertion.
• step description "@FairStep(fw) def mult(walrus, bird): """ Multiply two integers together (walrus and bird). """ result = walrus * bird return result " assertion.
• step description "@FairStep(fw) def add(dog, cat): """ Add two integers together (dog and cat). """ result = dog + cat return result " assertion.
• step description "@FairStep(fw) def mult(walrus, bird): """ Multiply two integers together (walrus and bird). """ result = walrus * bird return result " assertion.
• step description "@FairStep(fw) def add(dog, cat): """ Add two integers together (dog and cat). """ result = dog + cat return result " assertion.
• step description "@FairStep(fw) def mult(walrus, bird): """ Multiply two integers together (walrus and bird). """ result = walrus * bird return result " assertion.
• step description "@FairStep(fw) def add(dog, cat): """ Add two integers together (dog and cat). """ result = dog + cat return result " assertion.
• step description "@FairStep(fw) def add(dog, cat): """ Add two integers together (dog and cat). """ result = dog + cat return result " assertion.
• step description "@FairStep(fw) def mult(walrus, bird): """ Multiply two integers together (walrus and bird). """ result = walrus * bird return result " assertion.

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