Matches in Nanopublications for { ?s <http://purl.org/dc/elements/1.1/description> ?o ?g. }
- 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 "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 "Add 10 ml TE buffer into the mortar and carefully bring mycobacteria in suspension. Transfer this suspension to a 50 ml polypropylene tube." 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 "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 "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 "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 "Wash DNA pellet by adding 10 ml of 75% ethanol. Centrifuge at 12,000 x g for 10 min, 4 °C." assertion.
- step description "Centrifuge tube at 12,000 x g for 10 min, 4 °C. Discard liquid carefully." assertion.
- step 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)." 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 "Repeat steps 6 to 8 for achieving a higher purity of DNA." 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 "Add an equal volume of PCI mixture to the tube and mix contents by gently inverting tube for 5-10 min." 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 "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 "Pellet down bacilli by centrifugation at 2,000 x g for 10 min, room temperature and carefully discard supernatant." 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 "Add 10 ml TE buffer into the mortar and carefully bring mycobacteria in suspension. Transfer this suspension to a 50 ml polypropylene tube." assertion.
- step 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" 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 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 "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)" 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 "Dissolve pellet in molecular biology grade water and analyze DNA by agarose gel electrophoresis and spectrophotometer for its purity and integrity" assertion.
- step 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." 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 "Aspirate the whitish buffy coat (about 1 ml) (PBMCs) formed in the interphase between histopaque and medium." 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 "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." assertion.
- step description "Take 4 ml of Ficoll Histopaque in a 15 ml centrifuge tube." assertion.
- step description "Collect 4 ml of human venous blood sample in heparinised vials (BD biosciences) and mix well by gently inverting the tube several times." assertion.
- step 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." assertion.
- step 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." assertion.
- step 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." assertion.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.
- small_corpus.bel description "Approximately 2000 hand curated statements drawn from 57 PubMeds." provenance.