References for: description

Full identifier: http://purl.org/dc/elements/1.1/description

Nanopublication Part Subject Predicate Object Published By Published On
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
description
(unknown)
2024-04-17T04:18:57.584Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Project 02
(unknown)
2024-04-17T04:09:03.226Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Project 2 Description
(unknown)
2024-04-17T03:50:45.148Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Test
(unknown)
2024-04-17T03:35:43.982Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
The back yard at my house
(unknown)
2024-04-17T03:15:57.630Z
links a nanopublication to its pubinfo http://www.nanopub.org/nschema#hasPublicationInfo pubinfo
description
This set includes the nanopublications of the submissions, auxiliary class definitions, reviews, responses, and decisions for the special issue at the journal Data Science by IOS Press.
Cristina-Iulia Bucur
2022-03-01T10:59:05.050Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Decontaminate the tools for muscle dissection, including forceps, scalpels, and scissors, with RNaseZAPTM wipes and rinse thoroughly with double-distilled water (ddH2O). Decontaminate the procedure area by spraying with 70% ethanol.
Nanotate
2021-05-19T09:18:52.902Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Gently layer the blood on the top of Ficoll Histopaque using a 1 ml auto pipette. The layering should be done very slowly that blood and Ficoll Histopaque should stay as two different layers.
Nanotate
2021-03-23T16:38:19.974Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash (centrifuge in 100 x g for 10 min) twice with 10 ml of sterile PBS or sterile Dulbecco's modified eagle medium. The approximate yield of cells from 4 ml of blood varies between 107-108.
Nanotate
2021-03-23T16:35:38.985Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash (centrifuge in 100 x g for 10 min) twice with 10 ml of sterile PBS or sterile Dulbecco's modified eagle medium. The approximate yield of cells from 4 ml of blood varies between 107-108.
Nanotate
2021-03-23T16:30:58.156Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Collect 4 ml of human venous blood sample in heparinised vials (BD biosciences) and mix well by gently inverting the tube several times.
Nanotate
2021-03-22T16:51:24.240Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Take 4 ml of Ficoll Histopaque in a 15 ml centrifuge tube.
Nanotate
2021-03-22T16:51:11.919Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Gently layer the blood on the top of Ficoll Histopaque using a 1 ml auto pipette. The layering should be done very slowly that blood and Ficoll Histopaque should stay as two different layers.
Nanotate
2021-03-22T16:50:58.977Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-22T16:50:45.625Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Aspirate the whitish buffy coat (about 1 ml) (PBMCs) formed in the interphase between histopaque and medium.
Nanotate
2021-03-22T16:50:34.183Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-22T16:50:22.814Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash (centrifuge in 100 x g for 10 min) twice with 10 ml of sterile PBS or sterile Dulbecco's modified eagle medium. The approximate yield of cells from 4 ml of blood varies between 107-108.
Nanotate
2021-03-22T16:50:12.074Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Dissolve pellet in molecular biology grade water and analyze DNA by agarose gel electrophoresis and spectrophotometer for its purity and integrity
Nanotate
2021-03-22T16:35:18.770Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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
Nanotate
2021-03-22T16:32:27.978Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add an equal volume of isopropanol to tube. Mix contents gently and keep undisturbed for 5-10 min. The white thread-like structure of precipitated DNA will be seen in suspension (Figure 1)
Nanotate
2021-03-22T16:28:16.504Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge suspension at 12,000 x g for 10 min, 4 °C. Carefully transfer upper aqueous phase to a new 50 ml tube
Nanotate
2021-03-22T16:25:19.318Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-22T16:20:18.128Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add an equal volume of isopropanol to tube. Mix contents gently and keep undisturbed for 5-10 min. The white thread-like structure of precipitated DNA will be seen in suspension
Nanotate
2021-03-22T16:17:55.678Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add 10 ml TE buffer into the mortar and carefully bring mycobacteria in suspension. Transfer this suspension to a 50 ml polypropylene tube.
Nanotate
2021-03-22T16:13:19.826Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-22T16:10:30.726Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Pellet down bacilli by centrifugation at 2,000 x g for 10 min, room temperature and carefully discard supernatant.
Nanotate
2021-03-22T16:07:32.719Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-22T16:05:00.244Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-21T08:06:57.474Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add an equal volume of PCI mixture to the tube and mix contents by gently inverting tube for 5-10 min.
Nanotate
2021-03-20T05:42:29.104Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge suspension at 12,000 x g for 10 min, 4 °C. Carefully transfer upper aqueous phase to a new 50 ml tube.
Nanotate
2021-03-20T05:42:13.116Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Repeat steps 6 to 8 for achieving a higher purity of DNA.
Nanotate
2021-03-20T05:42:01.565Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add 100 µl 3 M sodium acetate solution per ml of aqueous phase obtained in step 10. Mix gently by inverting tube.
Nanotate
2021-03-20T05:41:45.421Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add an equal volume of isopropanol to tube. Mix contents gently and keep undisturbed for 5-10 min. The white thread-like structure of precipitated DNA will be seen in suspension (Figure 1).
Nanotate
2021-03-20T05:41:26.162Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge tube at 12,000 x g for 10 min, 4 °C. Discard liquid carefully.
Nanotate
2021-03-20T05:41:11.954Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash DNA pellet by adding 10 ml of 75% ethanol. Centrifuge at 12,000 x g for 10 min, 4 °C.
Nanotate
2021-03-20T05:40:57.160Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-20T05:40:42.587Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-20T05:40:20.117Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-20T05:40:04.034Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-20T05:39:47.164Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Pellet down bacilli by centrifugation at 2,000 x g for 10 min, room temperature and carefully discard supernatant.
Nanotate
2021-03-20T05:39:30.983Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add 10 ml TE buffer into the mortar and carefully bring mycobacteria in suspension. Transfer this suspension to a 50 ml polypropylene tube.
Nanotate
2021-03-20T05:38:47.140Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-20T05:38:29.271Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Dissolve pellet in molecular biology grade water and analyze DNA by agarose gel electrophoresis and spectrophotometer for its purity and integrity.
Nanotate
2021-03-20T05:37:37.743Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
At the indicated time points (0, 2, 4, 8, 12, 16, 20, 24, 28 h post-infection), cells were rinsed with 10 ml Phosphate Buffered Saline (PBS) buffer once and lysed in 1 ml TRIzol for 5 min at room temperature.
Nanotate
2021-03-19T15:27:20.144Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
RNA pellet was washed with 1 ml 70% RNase-free ethanol once and spin down by 7,500 x g for 5 min.
Nanotate
2021-03-19T15:22:02.652Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
RNA was precipitated by centrifugation at 12,000 x g for 10 min at 4 °C.
Nanotate
2021-03-19T15:10:58.508Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-19T15:07:42.065Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
RNA was precipitated by centrifugation at 12,000 x g for 10 min at 4 °C.
Nanotate
2021-03-19T15:01:25.318Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-18T16:08:37.156Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge the tube for 10 min at 5000 × g at 4°C. Transfer the upper aqueous phase into new 2ml tubes and add 5μl RNAse A (10 mg/mL). Place the tube for 30 min at 37°C.
Nanotate
2021-03-18T16:05:25.412Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Dissolve the pellet in 20-30 µl of RNase free water (commercial) or autoclaved water.
Nanotate
2021-03-13T15:51:14.285Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
After centrifuging at 5400 g at 4˚C for 5 min, remove the supernatant and then air-dry the pellet.
Nanotate
2021-03-13T15:48:48.317Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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
Nanotate
2021-03-13T15:45:52.777Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Transfer the upper aqueous phase to a new tube (size= 1.5 ml)
Nanotate
2021-03-13T15:42:40.695Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Dissolve the pellet in 20-30 µl of RNase free water (commercial) or autoclaved water.
Nanotate
2021-03-13T15:36:29.162Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
After centrifuging at 5400 g at 4˚C for 5 min, remove the supernatant and then air-dry the pellet.
Nanotate
2021-03-13T15:32:59.311Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-13T08:19:56.557Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-12T10:42:19.660Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-12T10:26:15.758Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge the tube for 10 min at 5000 × g at 4°C. Transfer the upper aqueous phase into new 2ml tubes and add 5μl RNAse A (10 mg/mL). Place the tube for 30 min at 37°C.
Nanotate
2021-03-11T16:43:03.902Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-11T16:42:19.939Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash DNA pellet by adding 10 ml of 75% ethanol. Centrifuge at 12,000 x g for 10 min, 4 °C
Nanotate
2021-03-10T11:23:07.795Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-10T11:20:30.406Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-10T11:19:16.882Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-10T11:18:25.462Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Pellet down bacilli by centrifugation at 2,000 x g for 10 min, room temperature and carefully discard supernatant.
Nanotate
2021-03-10T11:15:18.866Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-10T11:13:12.232Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge the tube for 10 min at 5000 × g at 4°C. Transfer the upper aqueous phase into new 2ml tubes and add 5μl RNAse A (10 mg/mL). Place the tube for 30 min at 37°C.
Nanotate
2021-03-10T10:53:40.132Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-10T10:17:44.515Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
RNA was precipitated by centrifugation at 12,000 x g for 10 min at 4 °C.
Nanotate
2021-03-05T15:58:44.249Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
RNA pellet was washed with 1 ml 70% RNase-free ethanol once and spin down by 7,500 x g for 5 min.
Nanotate
2021-03-05T15:58:00.648Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-05T11:37:30.479Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
After centrifuging at 5400 g at 4˚C for 5 min, remove the supernatant and then air-dry the pellet.
Nanotate
2021-03-05T11:36:37.574Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Dissolve the pellet in 20-30 µl of RNase free water (commercial) or autoclaved water.
Nanotate
2021-03-05T11:35:43.300Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Collect 4 ml of human venous blood sample in heparinised vials (BD biosciences) and mix well by gently inverting the tube several times.
Nanotate
2021-03-04T15:12:03.672Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Take 4 ml of Ficoll Histopaque in a 15 ml centrifuge tube.
Nanotate
2021-03-04T15:11:43.339Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Gently layer the blood on the top of Ficoll Histopaque using a 1 ml auto pipette. The layering should be done very slowly that blood and Ficoll Histopaque should stay as two different layers.
Nanotate
2021-03-04T15:11:14.099Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-04T15:10:39.161Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Aspirate the whitish buffy coat (about 1 ml) (PBMCs) formed in the interphase between histopaque and medium.
Nanotate
2021-03-04T15:10:07.828Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-04T15:09:22.152Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-02-26T09:51:16.417Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge suspension at 12,000 x g for 10 min, 4 °C. Carefully transfer upper aqueous phase to a new 50 ml tube.
Nanotate
2021-02-26T09:50:18.741Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Repeat steps 6 to 8 for achieving a higher purity of DNA.
Nanotate
2021-02-26T09:49:33.122Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add 100 µl 3 M sodium acetate solution per ml of aqueous phase obtained in step 10. Mix gently by inverting tube.
Nanotate
2021-02-26T09:49:12.595Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge tube at 12,000 x g for 10 min, 4 °C. Discard liquid carefully.
Nanotate
2021-02-26T09:48:41.071Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash DNA pellet by adding 10 ml of 75% ethanol. Centrifuge at 12,000 x g for 10 min, 4 °C.
Nanotate
2021-02-26T09:48:23.647Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-02-26T09:44:58.004Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-02-26T09:44:15.524Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add an equal volume of PCI mixture to the tube and mix contents by gently inverting tube for 5-10 min.
Nanotate
2021-02-26T09:40:50.359Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Dissolve pellet in molecular biology grade water and analyze DNA by agarose gel electrophoresis and spectrophotometer for its purity and integrity.
Nanotate
2021-02-26T09:39:38.488Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
At the indicated time points (0, 2, 4, 8, 12, 16, 20, 24, 28 h post-infection), cells were rinsed with 10 ml Phosphate Buffered Saline (PBS) buffer once and lysed in 1 ml TRIzol for 5 min at room temperature.
Nanotate
2021-02-18T09:45:01.698Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Collect 4 ml of human venous blood sample in heparinised vials (BD biosciences) and mix well by gently inverting the tube several times.
Nanotate
2021-02-14T04:57:52.681Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Take 4 ml of Ficoll Histopaque in a 15 ml centrifuge tube.
Nanotate
2021-02-14T04:57:27.221Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-02-14T04:56:14.950Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Aspirate the whitish buffy coat (about 1 ml) (PBMCs) formed in the interphase between histopaque and medium.
Nanotate
2021-02-14T04:55:43.561Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-02-14T04:55:00.251Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
has the description
Tobias Kuhn
2020-12-18T09:33:57.621Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add imaging buffer with desired ratios of Buffer C (500 mM), ethylene carbonate, and IS-CF660R at 1-2 nM final concentration. The exact concentration of IS may need to be adjusted depending on the target and based on the imaging kinetics.
(unknown)
2020-11-14T18:05:21.158Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add imaging buffer with desired ratios of Buffer C (500 mM), ethylene carbonate, and IS-CF660R at 1-2 nM final concentration. The exact concentration of IS may need to be adjusted depending on the target and based on the imaging kinetics.
(unknown)
2020-09-27T00:50:41.265Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Add imaging buffer with desired ratios of Buffer C (500 mM), ethylene carbonate, and IS-CF660R at 1-2 nM final concentration. The exact concentration of IS may need to be adjusted depending on the target and based on the imaging kinetics.
(unknown)
2020-09-27T00:50:41.265Z