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
(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
Add 500 μl phenol:chloroform:isoamyl alcohol (25:24:1) to the tube. Shake vigorously to mix thoroughly. Centrifuge the samples at 12,000 x g for 15 min.
Nanotate
2022-01-11T06:17:07.203Z
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.
Nanotate
2022-01-11T06:08:46.582Z
links a nanopublication to its pubinfo http://www.nanopub.org/nschema#hasPublicationInfo pubinfo
description
A dataset of 327 off-label drug indications found in PubMed articles. With additional information on the context of the indications, such as the target group age range (adult/children), or if the target group has a specific phenotype. Drugs are identified by their DrugBank IDs, and conditions are identified by their MONDO, EFO, or HPO IDs. Curated by Ricardo de Miranda Azevedo. See https://github.com/MaastrichtU-IDS/off-label-drug-indications-dataset for more details.
Vincent Emonet
2021-10-02T00:00:00Z
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
The RNA pellets are air-dried and dissolved in 100 µl RNase-free H2O by incubating at 65 °C for 15 min.
Nanotate
2021-03-19T15:16:44.868Z
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
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.
Nanotate
2021-03-19T14:56:48.604Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Cell lysates were transfer into eppendorf tubes and one-fifth (volume/volume) of chloroform was added to each tube.
Nanotate
2021-03-19T14:53:59.410Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
The IBV-infected cells were incubated at 37 °C in 5% CO2
Nanotate
2021-03-19T14:52:04.142Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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).
Nanotate
2021-03-18T16:15:11.750Z
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
2021-03-18T16:12:45.992Z
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 dry pellet in nuclease free water or TE buffer (Ph 8.0). Store DNA at 4 º celcius or -20 º celcius .
Nanotate
2021-03-18T06:34:27.555Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Keep the pellet at 37 º celcius for 10 minutes.
Nanotate
2021-03-18T06:32:53.163Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash the pellet with 70% ethanol .
Nanotate
2021-03-18T06:31:23.679Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash preserved mosquitoes in sterile distilled water or phosphate buffer saline(PBS) to remove excess alcohol. Fresh mosquitoes can be ground directly.
Nanotate
2021-03-18T06:28:01.670Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-16T16:48:04.108Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Keep the pellet at 37 º celcius for 10 minutes
Nanotate
2021-03-16T16:42:33.652Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash the pellet with 70% ethanol
Nanotate
2021-03-16T16:38:16.957Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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
Nanotate
2021-03-16T16:33:42.128Z
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
Cell lysates were transfer into eppendorf tubes and one-fifth (volume/volume) of chloroform was added to each tube
Nanotate
2021-03-11T17:36:57.258Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
RNA concentration and purity were determined by NanoDrop.
Nanotate
2021-03-11T17:31:58.912Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
The IBV-infected cells were incubated at 37 °C in 5% CO2.
Nanotate
2021-03-11T17:15:37.640Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-11T16:45:43.287Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-11T16:45:05.510Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
The falcon tube was kept into the 65°C incubator or water bath and mix gently by inversion after every 10 min till 45 min. After incubation, place the tube at room temperature for five min to reach to room temperature environment. Centrifuge the 50ml falcon tube for 5 min at 3000×g on room temperature. For next generation sequencing, the greater the genome size, lower is speed of initial centrifugation
Nanotate
2021-03-11T16:44:41.382Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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.
Nanotate
2021-03-11T16:43:40.490Z
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
Add 2 volumes of cold 95% ethanol and mix gently by inversion. The tubes are incubated at for 45 min in -20°C freezer. It should not be for more time as some remaining phenolics and resin may also precipitate with DNA.
Nanotate
2021-03-11T16:41:02.775Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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).
Nanotate
2021-03-11T16:39:49.461Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Lyse for 1 hour 30 minutes at 37º celcius. Gently mix the tube by inverting every 15 minutes.
Nanotate
2021-03-11T10:51:04.012Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge at 12,000g for 10 minutes at 4º celcius .Transfer the supernatant to a fresh tube.
Nanotate
2021-03-11T10:50:44.677Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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 .
Nanotate
2021-03-11T10:50:24.056Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Repeat the above step(5) , then add chloroform:isoamyl alcohol(24:1) and centrifuge at 12,000g for 10 minutes at 4º celcius .
Nanotate
2021-03-11T10:49:52.989Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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 .
Nanotate
2021-03-11T10:49:31.220Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Centrifuge at 12,000g for 30 minutes at 4º celcius and then remove the supernatant .
Nanotate
2021-03-11T10:49:02.879Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash the pellet with 70% ethanol .
Nanotate
2021-03-11T10:48:46.263Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Keep the pellet at 37 º celcius for 10 minutes.
Nanotate
2021-03-11T10:48:26.760Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
description
Wash preserved mosquitoes in sterile distilled water or phosphate buffer saline(PBS) to remove excess alcohol. Fresh mosquitoes can be ground directly.
Nanotate
2021-03-11T10:48:09.499Z
links a nanopublication to its assertion http://www.nanopub.org/nschema#hasAssertion assertion
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 .
Nanotate
2021-03-11T10:45:27.539Z
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
2021-03-10T11:30:16.993Z
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
2021-03-10T11:27:29.159Z
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