Tubes washing

Pass 0.5 mL of the suspended acetone solution through the Bond Elut SCX (100-mg) cartridge and collect in an Eppendorf tube. Wash the cartridge column with two portions of 0.5 mL of acetone and combine all of the eluates (total volume ca 1.5 mL). This step is omitted for water. 

Scrape the cells from each flask and transfer them into a 50 ml tube. Wash each flask using a single 10 ml portion of 0.025 M Tris, 0.15 M NaCl, pH 7.2, and add the solution to the scraped cells. 

After the combustion is finished wash out the tube as described for the halogen determination, collecting the liquid in a small clean beaker and add the contents of the test tube, washing it out also. Then add a clear filtered mixture of 2-3-5 c.c. of barium chloride solution (1 10) and 10 drops of dilute hydrochloric acid, cover with a clean watch-glass, heat to boiling, and continue until separation of barium sulphate ceases. Immerse the beaker in cold water to cool and complete the determination as described in the preceding section. 

The solution is cooled to -78°C in a dry ice-acetone bath, and 38.19 g (0.23 mol) of tetrafluoroboric acid-diethyl ether complex (Note 10) is added dropwise by syringe over a 30-min period. The solution is allowed to warm to room temperature. The yellow precipitate is filtered off and collected in a 250-mL Schlenk tube, washed with ether, dried under a stream of nitrogen and finally under reduced pressure (oil pump). The bright yellow salt 1 weighs 43. 2-57. 6 g (60-80%) and may be used without further purification (Note 11). U may be stored indefinitely under nitrogen at 0 C without decomposition. 

A more socially acceptable option, is to water-wash the tubes. Most of the effective washing must be underneath the tubes. Washing from the top down is relatively ineffective. In many cases, detergent must be added to the washwater to remove greasy dirt. (Caution Hot tubes may be thermally shocked by this washing, and pull out of the tube header box.)... 

A warm (60°C) solution of sodium perrhenate (1.5 g, 5.5 mmol) in demineralized water (60 mL) in a 250-mL conical flask is added to a warm (60°C) solution of tetraphenylphosphonium bromide (2.8 g, 6.7 mmol) in demineralized water (60 mL) contained in a 250-mL conical flask. The mixture is then stirred for 30 min at room temperature. The white precipitate which forms is collected by filtration on a No. 3 sintered-glass filter tube, washed with distilled water (4 x 20 mL) and absolute ethanol (4 x 30 mL), and then dried in vacuo for 5 h at 56°C (boiling acetone) (yield 2.80 g, 87%). 

Into a test-tube, washed out with boiled water, are poured 10 c.c. of the unfiltered honey solution and exactly 1 c.c. of a fresh, clear 1 % solution of soluble starch.8 After thorough mixing, the tube is immersed in a water-bath at 450 for exactly an hour. A few drops of iodine solution (1 gram of iodine and 2 grams of potassium iodide in 200 c.c. of water) are then added and the colour at once assumed by the liquid observed. 

Transfer 100 pg of MB into 0.5 mL Eppendorf tube. Wash the MB once with 100 pL of TTL buffer and then separate, decant and... 

Filter the aqueous layer from Step 5 through Whatman paper and collect the effluent in a glass tube. Wash the original tube and filter 2-3 times with 5 ml methanol, then dry the filtrate with a nitrogen stream. 

Transfer small amount of resin to the test tube, wash resin with DCM, and perform the bromophenol blue (BPB) test (29) for presence of free amine (see Note 6). 

Step 9. Dissolve the Pdl2 in 5 mL of 6 M NH4OH and heat in a boiling water bath for about 5 minutes. If any residue remains, centrifuge and decant the supernatant into a clean centrifuge tube. Wash the residue with 5 ml water, centrifuge, and add wash to the supernatant solution in the other centrifuge tube. 

The resultant cell suspension was then transferred into 15-mL Falcon tubes, washed with IX PBS or RPMI, and the cells were counted. 

Tubing wash. Removes rust, mill scale and precipitates 15% HC1 with inhibitor is circulated in the tubing followed backwashing from the well. 

At the conclusion of the experiment the sublimate was carefully removed from the cold finger and weighed. The weight of the residue was found by weighing the outer tube, washing out the residue, and reweighing the tube. The analyses were performed by iodometry. 

Number the vials and record their contents. Use leaves nearest the growing point of a single stem. Do not include flowers or flower buds Use the glass rod to push the leaves to the bottom of the tube. Wash and dry the rod between tests. 

Carefully layer the Percoll solutions sequentially into a test tube. Wash cells in PBS, resuspend at a high concentration (typically 20 x 10 mL ) in PBS and place on top of the Percoll column. 

Harvest cells and count using a hemocytometer (see Note 2). Place aliquots of 1 x 106 cells into flow cytometer tubes. Wash once in ice-cold PBS and pour off the PBS, leaving the pellet with a small amount of fluid. Cells are pelleted at... 

Transfer cells to a universal tube, wash twice with media containing 1 mM glutamine and 10% FCS maintained at 4°C... 

PROBE WASH STATION /— BEAD/TUBE WASH STATION... 

Resuspend the clear RNA pellet in 200 pi autodigestedd proteinase K solution and transfer to a microfuge tube. Wash the polyallomer tube bottom with an additional 200 pi of proteinase K solution and combine in the microfuge tube. 

For preparation of ethyl isobutyl ether,724 sodium (9 g) is dissolved in pure isobutyl alcohol, and the excess of alcohol is distilled off in a stream of hydrogen at 180°. The residual sodium isobutoxide is rapidly powdered and then heated in a closed tube at 75° with ethyl iodide (60 g) until all the iodide has reacted (about 7 days). Isobutene escapes when the tube is opened. The ethyl isobutyl ether formed is distilled from the tube, washed with water, dried, and finally distilled over sodium (b.p. 79-80°). The yield is good. 

After stirring for an additional hour, the solid is collected by filtration through a Schlenk filter tube, washed with 50 mL of dry benzene and 50 mL of dry diethyl ether, and dried under vacuum. Benzene and diethyl ether should be heated at reflux over sodium metal. The yield of crude [PhjI —CCljjCl is approximately 40 g. The product must be recrystallized before it is used as a starting material for Part B. 

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