Fluid transfer control for real-time pcr

A method for controlling transfer of a solution to a vessel for subsequent real-time PCR amplification in the vessel, the method comprising the steps of:

• providing a solution comprising a concentration of a control dye, wherein the solution also comprises a detection dye for the PCR amplification,
• transferring a pre-defined volume of the solution comprising the control dye to each of a plurality of vessels,
• measuring an intensity of the control dye within each vessel,
• comparing the intensity measured for each vessel with a pre-defined threshold value, whereby a fluid transfer error is detected for a vessel if the measured intensity is below the pre-defined threshold value,
• compiling a mean intensity value of all vessels having a measured intensity that is above the pre-defined threshold value,
• verifying for each vessel having a measured intensity that is above the pre-defined threshold value, if the intensity is within a pre-defined range surrounding the compiled mean intensity value, wherein a fluid transfer error is detected for a vessel if the measured intensity is not within the pre-defined range,
• declaring a correct fluid transfer for each vessel having a measured intensity that is within the pre-defined range, and
• performing the PCR amplification in the plurality of vessels and marking accordingly results of the amplifications performed in vessels wherein an incorrect fluid transfer is declared, or performing PCR amplification in vessels wherein correct fluid transfer is declared.

A method for controlling transfer of a solution to a vessel for subsequent real-time PCR amplification in the vessel, the method comprising the steps of:

• providing a solution comprising a concentration of a control dye, wherein the solution also comprises a detection dye for the PCR amplification,
• transferring a pre-defined volume of the solution comprising the control dye to each of a plurality of vessels,
• measuring an intensity of the control dye within each vessel,
• comparing the intensity measured for each vessel with a pre-defined threshold value, whereby a fluid transfer error is detected for a vessel if the measured intensity is below the pre-defined threshold value,
• compiling a mean intensity value of all vessels having a measured intensity that is above the pre-defined threshold value,
• verifying for each vessel having a measured intensity that is above the pre-defined threshold value, if the intensity is within a pre-defined range surrounding the compiled mean intensity value, wherein a fluid transfer error is detected for a vessel if the measured intensity is not within the pre-defined range,
• declaring a correct fluid transfer for each vessel having a measured intensity that is within the pre-defined range,
• transferring a pre-defined volume of a second solution comprising a concentration of a quencher molecule to each of the vessels, wherein the quencher molecule quenches the intensity of the control dye,
• measuring a quenched intensity of the control dye within each vessel,
• calculating a quenching parameter value based on the measured quenched intensity for each vessel, wherein a fluid transfer error for a vessel is detected if a pre-defined quenching threshold criterion of the quenching parameter value is not fulfilled,
• compiling a mean quenching parameter value of all vessels having a calculated quenching parameter value that fulfills the pre-defined quenching threshold criterion,
• verifying for each vessel having a calculated quenching parameter value that fulfills the pre-defined quenching threshold criterion, if the quenching parameter value is within a pre-defined second range surrounding the compiled mean quenching parameter value, whereby a fluid transfer error is detected for a vessel, if the measured quenching parameter value is not within the pre-defined second range,
• declaring a second correct fluid transfer for each vessel having a measured quenching parameter value that is within the pre-defined second range, and
• performing the PCR amplification in the plurality of vessels and marking accordingly results of the amplifications performed in vessels wherein an incorrect fluid transfer is declared, or performing PCR amplification in vessels wherein correct fluid transfer is declared.