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#Serial dilution calculator how to
#Serial dilution calculator serial
#Serial dilution calculator series

In this example, although the high-efficiency condition (the blue curve in Figure 5) gives a later C t at high concentrations, it results in better sensitivity at low target concentrations. In Figure 5, two samples (X and Y) amplified under low and high efficiency conditions show different C t values for the same target concentration.

#Serial dilution calculator series

A dilution series amplified under low efficiency conditions could yield a standard curve with a different slope from one amplified under high efficiency conditions. The efficiency of a PCR reaction can also affect C t. Therefore, the C t values from PCR reactions run under different conditions or with different reagents cannot be compared directly. However, artifacts from the reaction mix or instrument that change the fluorescence measurements associated with the C t calculation will result in template-independent changes to the C t value. The C t value increases with a decreasing amount of template. The threshold must be set in the linear phase of the amplification plot in Figure 1C.

The exponential phase in Figure 1B corresponds to the linear phase in Figure 1C.

#Serial dilution calculator how to

We will discuss the most common template-independent factors that can influence C t and describe how to evaluate the performance of a real-time PCR reaction.įigure 1, above, shows several parameters of the real-time reaction amplification plot. Many factors impact the absolute value of C t besides the concentration of the target. It is a relative measure of the concentration of target in the PCR reaction.

As six tubes are used, the final reduction of germs/cells will be 10 -6 (1 in 1,000,000).C t (threshold cycle) is the intersection between an amplification curve and a threshold line ( Figure 1B).

Repeat the same process with the remaining test tube, taking 1 ml from the previous tube and adding to the next 9 ml diluents.

The second tube now has a total dilution factor of 10 -2.

Then dilute it and pour into a second test tube.

Now, take 1 ml of mixture from the 10 -1 concentration test tube.

Discharge a tip of pipette, and insert a new pipe tip into the pipette.

Dilution is thoroughly mixed by removing and filling the pipette several times.

This provides an initial dilution of 10 -1.

Then add sample solution to the first tube to form a total volume of 10 ml.

Add 1 ml of a well-mixed sample/culture is drawn into the pipette.Take a given sample/culture in a test tube and six test tubes, each containing 9 ml of sterile diluent, which can be pure water or 0.9% saline.

#Serial dilution calculator serial

The serial dilution formula for the dilution factor of each tube is –īeneath is an elucidated ten-step process designed to achieve a purification factor of 10 -6, providing an extensive understanding of every stage involved.The dilution factor in serial dilution can either be ascertained per test tube or taken into account as a collective diluting entity throughout the entire dilution series.Serial two-fold and ten-fold dilutions are commonly used to titer antibodies or prepare diluted analytes in the laboratory.On the other hand, for a less contaminated sample, a low dilution factor might be sufficient. For example, if a water sample is taken from a very polluted environment, the dilution factor is increased. Depending on the relative concentration of cells/organisms in the sample, the size of the stretch is determined.After that, a small average volume of each purifier is used to make a series of pouring or dispersing plates.Serial dilution involves the process of taking a sample and diluting it through a series of standard volumes of sterile diluent, which can be either distilled water or 0.9 % saline solution.