Sustainable Alternatives To Paper Towels
Place tissue on a flat surface (dissecting board) covered with dry paper towels and remove fat, necrotic tissue and gross debris (see Fig. 2). 2. Place slightly stretched tissue flat on paper towels, mucosal side up. Using curved fine forceps, gently pinch and lift the mucosa at one edge of the specimen. Cut between the mucosal and the muscle layers with fine curved iris scissors, starting at the lifted edge of the specimen and if possible, longitudinally to the circular
Windowing the egg Eggs should be cleaned (but not soaked) with 70 ethanol on a damp paper towel. Puncture the air space at the blunt end of the egg by piercing the shell with a needle. After air has escaped, the hole will be sealed by the shell membranes. With the egg's long axis parallel to the horizon, window the egg shell by carefully removing, using a dental drill and forceps, approx 1 cm2 of egg shell and the underlying shell membranes at the point marked on the side of the egg (i.e., the point that was uppermost during incubation).
The Basic Protocol produces gels with separated polypeptides that can be subjected to standard electrotransfer procedures (unit 10.7). It is necessary, however, to remove the nonionic detergent (included in the IEF gel recipe) from the gel as it interferes with a number of commonly used staining protocols the nonionic detergent also very significantly reduces the amount of protein transferred to membranes in electroblotting experiments. It is also necessary to bind SDS to the proteins. Both tasks are accomplished by repeated washings of the gel as follows The gel is submerged in a shallow dish containing 50 methanol 1 (w v) SDS 5 mM TrisCl (pH 8.0) the volume of wash solution should be 10x the volume of the gel. The gel is washed five times, 10 min each time, changing the solutions between washes. A convenient way to restrain the gel while decanting the used wash solution is to place paper towels or filter paper of a size that approximates that of the gel directly on the surface of...
One disadvantage with the upward capillary method (basic protocol) is that the gel can become crushed by the weighted filter papers and paper towels that are laid on top of it, reducing capillary action. The problem worsens as the transfer proceeds, as the paper towels become soaked in transfer buffer, increasing the pressure on the gel. This retards the blotting process and as a result the transfer must be carried out for 16 to 24 hr. 2. Make a stack of paper towels 2 to 3 cm high in a glass dish. The towels should be slightly wider than the gel. 3. Using Figure 10.6.2 as a guide, place four pieces of Whatman 3MM filter paper on top of the paper towels. Wet a fifth filter paper with transfer buffer and place on top. membrane Whatman 3MM paper towels (2-3 cm) membrane Whatman 3MM paper towels (2-3 cm)
Anesthetize male rats by placing them in a bell jar containing ether. Once the rats are sedated, remove them from the bell jar and place them on their backs on a square sheet of bench protector in a fume hood. Place a 50-mL Falcon tube containing a piece of ether-soaked paper towel over the rat's nose to keep it sedated.
A positively charged nylon membrane does not have to be prewetted, but can be placed directly onto the gel. If uncharged nylon is being used, use 0.25 M NaOH 1.5 M NaCl as the transfer solution. Check that the paper towels are resistant to the alkali solution some types go brown and should not be used. 4. Remove the paper towels and filter paper and recover the membrane. Rinse the membrane in 2x SSC, place on a sheet of Whatman 3MM filter paper, and allow to air dry. Store as described in step 17 of the basic protocol.
BASIC Collection Counting and Culturing of Nippostrongylus brasiliensis Eggs and Protocol 2 Larvae from Infected Rats
In this protocol, infected rats are placed on wire-grid cages so fecal pellets fall through the grid onto paper towels which have been wetted sufficiently to remain moist (but not soaking) overnight. Collection starts on day 7 post-inoculation, with daily collections through to day 10 eggs on day 6 can be omitted for routine purposes, as egg numbers are low and variable.
Absorbent material (e.g., paper towels, Kimwipes) High-efficiency particulate air (HEPA) vacuum (Fisher) 4. Mop up the liquid with paper towels. Squeeze the solution out of the towels and collect in a suitable container. Discard towels as hazardous solid waste. 9. Verify complete decontamination by wiping the surface with a paper towel moistened with water and squeezing the liquid out of the towel. Test the liquid for the presence of benzidine or diaminobenzidine (see Support Protocol 1). Repeat steps 1 to 9 as necessary.
Wash the equipment contaminated with ethidium bromide once with a paper towel soaked in decontamination solution. The pH of the decontamination solution is 1.8. If this would be too corrosive for the surface to be decontaminated, wash with a paper towel soaked in water instead. 2. Wash the surface five times with paper towels soaked in water using a fresh towel each time. 6. Discard the decontamination solution and the paper towels as nonhazardous liquid and solid wastes.
Membrane described in the first basic and alternate protocols has certain disadvantages, notably the fact that the gel can become crushed by the weighted filter papers and paper towels that are laid on top of it. This slows down the blotting process and may reduce the amount of DNA that can be transferred. The downward capillary method described in the second alternate protocol is therefore more rapid than the basic protocol and can result in more complete transfer.
The weight should be sufficient to compress the paper towels to ensure good contact throughout the stack. Excessive weight, however, will crush the gel and retard transfer. 21. Remove paper towels and filter papers and recover the membrane and flattened gel. Mark the position of the wells on the membrane in pencil and ensure that the up-down and back-front orientations are recognizable.
In a Class II biohazard hood, place the animal on its back on clean, dry, absorbent paper towels covering a dissection board. 15. With hands in glove ports, carefully remove foam stoppers from the homogenization tubes and place them on a paper towel located within the glove box chamber. Carefully open the pestle packs and place the chilled pestles into the homogenization tubes, taking care not to splash or spill the tube contents.
Surgery should be done in a clean environment that has little to no traffic and away from heating or cooling vents (i.e., no major air flow see Note 6). Place the mouse in the lower level of the operation stage, on a few layers of paper towels so that the surface of the abdomen reaches the upper level of the stage. Ensure that the mouse is properly anesthetized by checking for a response after pinching the tail with forceps. Using microdissection scissors, make a mid-ventral incision of the skin (2- to 3-cm long), and then make a similar incision of the peritoneum (see Note 7). Use blunt forceps when manipulating the skin and fine forceps when manipulating the peritoneum. Retract both the skin and the peritoneum using S-shaped hooks attached to rubber bands tacked to the upper level of the operating stage.
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