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How much of each ingredient do I need to make a 1mL lysis buffer solution?
Wiki User
∙ 8y ago
You need to specify, lysis buffer for bacteria or eukaryotic cell. The most common buffer cocktail for eukaryotic cells is composed by an hypertonic solution and cell disrupters. To answer this question the proper amounts of the ingredients for 1 mL would be:
8.77 mg of NaCl (150 mM, the hypertonic component)
10 microliters NP-40 [can be replaced by 1 microliter (0.1%) of Triton X-100, a detergent]
6.06 mg of Tris in 100 microliters [Tris hydroxymethyl aminomethane, make 10 mL of a 100X solution (that is, 605.5 mg in 10 mL water, adjust to pH 8.0)]
890 microliters of distilled water.
In some cases can be added some protease inhibitors such as PMSF, leupeptin, Aprotinin, etc. at concentrations of 1 microgram/mL. Store at 4oC for one month.
Now, to make a lysis buffer for bacteria, the composition is different. For 1 mL mix:
6.06 mg of Tris in 100 microliters, as before
17.53 mg of NaCl (300 mM)
100 microliters of PMSF [Phenylmethylsulfonyl Fluoride, make 10X solution (1.742 mg in 10 mL of water)]
980 microliters of distilled water
add aprotinin, leupeptin, and pepstatin at final concentration of 1 microgram/mL.
Usually, to break the bacteria cell wall it is useful a lab blender. Disruption of both, eukaryotic and bacteria cells, must be done in cold conditions, usually on ice-water bath.
Wiki User
∙ 8y ago
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Does changing the amount of each ingredient change how the slime turns out?
yes
What is a eg of a solution?
two e.g of each group solution suspension and colloids and the names for each group
What are ion that conduct electricity in solution?
A solution containing ions conduct electricity. They are different for each solution.
What is the pH range of neutral solution?
The pKa of HOCl is 7.46. So a buffer of equal concentrations of each of HOCl and NaOCl will have a pH = 7.46, and so is slightly basic. However, the range of any buffer solution depends on the total concentration of the pair in the solution -- the more HOCl and NaOCl add the larger the range.
Why use different buffers for a range of ph?
Each buffer will only be of sufficient capacity within a pH interval of pKa+1 > pH > pKa-1 so the optimal buffer pH-range is maximum 2 units.
How can you describe the strength of a solution?
A solution is usually expressed as percentages of each constituent ingredient.
What are pH electrode storage solution and pH 7.0 buffer and for what are they used?
The buffer solution is used to calibrate the meter. Usually two buffer solutions are used to do this. One is close to the range of the sample that will be measured, like the 7.0 buffer that you mention could be used for aquarium water. The other solution would be more acidic, like a 4.0 buffer solution. The process is to calibrate in one solution, rinse the probe, then calibrate in the other solution. This back and forth is done until the meter is reading the desired buffer and is stable. In other words, you would finish with the 7.0 buffer solution when it is reading exactly 7.0 PH, with no further adjustments after switching from the 4.0 solution. The probe should be rinsed with PH neutral DI water and shaken dry between each between each measurement so that solutions are not mixed. The storage solution is used to store the glass probe for long periods of time. A few drops of storage solution should be dripped into the probe cap before replacing it for storage. The storage solution is just a buffer that has some anti-bacterial additives to keep stuff from growing in it. The calibration buffer can be used for short term storage though. The glass probe should NEVER be allowed to dry out for long periods and should be rinsed with DI water before storage. It should NOT be stored in DI water though. Only buffer solution or storage solution should be used.
How do you make 50 mM potassium phosphate buffer pH 7.4?
1 M Sodium Phosphate Buffer Stock Solution (1 liter) Protocol # Solution A: Dissolve 138.0 g NaH2PO4?H2O in 1 liter dH2O (pH 7.0). # Solution B: Dissolve 142.0 g Na2HPO4 in 1 liter dH2O (pH 7.0). # Mix 423 ml Solution A with 577 ml Solution B. # Autoclave and store at room temperature.
What are requirements for a buffer solution?
Requirements for a Buffer Solution:There are three requirements for buffer:• Must be a mixture of weak acid and its salt or weak base and its salt• A buffer must contain relatively large concentration of acid to react with added base (OH-) and also must contain similar concentration of base to reaction with added acid (H+).• The acid and base components of the buffer must not consume each other in a neutralization reaction.
What does buffer do?
A buffer prevents two things from touching or meeting, such as the buffer between two people keeps them from meeting and potentially harming each other.
What happens with a stong acid is added to a base?
The acid and base will neutralize each other and the resulting solution will be more neutral. If the base being neutralized is strong, the resulting salt will be neutral. If the base is weak the resulting salt will be acidic. A solution of such a salt may be called a buffer.
What has a definite amount of each ingredient?
compund
What is depth buffer method?
The Z-buffer algorithm is a convenient algorithm for rendering images properly according to depth. To begin with, a buffer containing the closest depth at each pixel location is created parallel to the image buffer. Each location in this depth buffer is initialized to negative infinity. Now, the zIntersect and dzPerScanline fields are added to each edge record in the polyfill algorithm. For each point that is to be rendered, the depth of the point against the depth of the point at the desired pixel location. If the point depth is greater than the depth at the current pixel, the pixel is colored with the new color and the depth buffer is updated. Otherwise, the point is not rendered because it is behind another object.
What is difference between single double and circular buffer?
single buffer : you read and write on the same buffer, can be messy if both reading and writing take place at the same time.double buffer : you read one buffer and you write the other one. When both reading and writing are complete, the buffers are swapped. It solves the problem of simultaneous reading and writing but requires synchronization.circular buffer : this a buffer with two pointer : read and write. If both pointers are equal, the buffer is empty.For each write operation, the write pointer advances and each time data is read back, the read pointer advances. It is circular because when a pointer reaches the end, it wraps back to the beginning.It may be used to implement a queue which allows simultaneous reading and writing without synchronization as long as the buffer is not full.A double buffer is basically a circular buffer of size 2 and a single buffer is basically a circular buffer of size 1.
What is the Difference between frame buffer and z buffer?
The z-buffer holds information about the depth of a 3D scene. Specifically, it holds information about the coordinates of each object in the scene, so that the CPU/GPU knows the order in which objects should be drawn.The frame buffer is a more physical thing. Your computer monitor stores information for the next image it's going to draw in the frame buffer.
How do buffers reduce changes in pH when hydrogen ions or hydroxide ions are added to a solution?
Buffer solutions act as neutralizing agents that contain both positive and negative ions of a weak acid. Thus, if a base such as NaOH is added to the solution, the positive H+ ions will react with the negative (OH-) ions to neutralize the solution. Similarly, if an acid is added to the buffer, the negative ions (such as OH- or C2H3O2-) will react with the hydronium ions to neutralize the solution.
How does changing the amount of each ingredient change how the slime turns out?
yes
