IP Library Granted Patent US 12,065,698
Granted Patent B2
US 12,065,698 · App. 17/498,308 · Granted Aug 20, 2024

Sample processing apparatus and method

Inventors: Jukka Tuunanen (Vantaa, FI); Jaako Kurkela (Vantaa, FI); Katja Eklin (Vantaa, FI)
Assignee: Thermo Fisher Scientific Baltics UAB
C12Q1/6876C12Q1/6846C12Q1/686G01N21/251G01N21/29G01N21/6428G01N21/78G01N2021/6439
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Quick Facts
Patent No.
US 12,065,698
App. No.
17/498,308
Granted
Aug 20, 2024
Kind
B2
Abstract

The invention relates to a sample processing apparatus comprising a holder for a microtiter plate comprising a plurality of microwells, optical measurement unit for measuring optical responses of samples dosed to the microwells, and a computing unit configured to analyze the optical responses in order to detect dosing failures in said plurality of microwells, and if a dosing failure has been detected in one or more of the microwells, to communicate the existence of the dosing failure to a user of the apparatus through signaling means or to store data indicative of the dosing failure to data storage means for further use. In particular, the invention relates to detecting dosing failures in before, during and after a PCR process.

Claims (36)

1. A method of preparing a reaction mixture for Polymerase Chain Reaction (PCR) assay, comprising

providing a first reagent solution comprising at least one substance required for performing said assay,

providing a second reagent solution comprising at least one other substance required for performing said assay,

mixing the first and second reagent solutions,

providing a third reagent solution comprising at least one further substance required for performing said assay,

mixing the third reagent solution with the first and second reagent solutions

for providing a mixed solution to be subjected to the PCR process, characterized in that

the first reagent solution contains a first colorant providing the first reagent solution a first color,

the second reagent solution contains a second colorant providing the solution a second color different from the first color,

said mixing of the first and second reagent solutions yields a mixed solution having, due to said first and second colorants, a third color different from the first and second colors, and wherein the third color is detected visually prior to subjecting the mixed solution to the PCR process,

the third reagent solution contains a third colorant providing the solution a fourth color different from the first, second, and third colors,

said mixing of the first, second, and third reagent solutions yields a mixed solution having, due to said first, second, and third colorants, a fifth color, different from the first, second, third, and fourth colors,

the first or second reagent solution comprises an Alexa 350, FAM, JOE, VIC, HEX, Cy3, TAMRA, Cy3.5, Texas Red, ROX, Cy5, Cy5.5, Quasar705, or SYBR dye.

2. A solution set for Polymerase Chain Reaction (PCR) assay, comprising

a first reagent solution comprising at least one substance required for performing said PCR assay,

a second reagent solution comprising at least one other substance required for performing said PCR assay,

a third reagent solution comprising at least one further substance required for performing said assay,

characterized in that

the first reagent solution is provided with a first colorant having a first color,

the second reagent solution is provided with a second colorant having a second color different from the first color,

the first and second reagent solutions are capable of forming, on mixing, a first mixed solution having, due to said first and second colorants, a third color different from the first and second colors, wherein the colors are capable of being detected visually,

the third reagent solution contains a third colorant providing the solution a fourth color different from the first, second, and third colors,

the first, second, and third reagent solutions are capable of forming, on mixing, a second mixed solution having, due to said first, second, and third colorants, a fifth color different from the first, second, third, and fourth colors,

the first or second reagent solution comprises an Alexa 350, FAM, JOE, VIC, HEX, Cy3, TAMRA, Cy3.5, Texas Red, ROX, Cy5, Cy5.5, Quasar705, or SYBR dye.

3. The solution set according to claim 2 , characterized in that at least one of said reagent solutions is a sample solution for receiving a template to be amplified.

4. The solution set according to claim 2 , characterized in that at least one of the reagent solutions contains one or more of the following: polymerase, primer, ions or dNTPs or fluorescent qPCR dye or probe, preferably being a PCR master mix, in particular a qPCR master mix.

5. The solution set according to claim 2 , characterized in that at least one of the reagent solutions is an elution buffer or a dilution buffer.

6. The solution set according to claim 2 , characterized in that said first and/or second colorants are selected from the group of: Quinoline yellow, Xylene cyanol, Brilliant Blue, Patent Blue, Indigocarmine, Acid Red 1, m-Cresol Purple, Cresol Red, Neutral Red, Bromocresol Green, Acid Violet 5, Bromo phenol blue, and Orange G.

7. The solution set according to claim 2 , characterized in that the concentration of said colorants in respective reagent solutions corresponds to absorbances of the said solutions at their maximum absorption wavelengths of 0.01-0.5, preferably 0.03-0.15 with 1 mm light path, when diluted to the desired PCR processing concentration.

8. The solution set according to claim 2 , characterized in that the first reagent solution and/or the second reagent solution is a concentrate.

9. The solution set according to claim 2 , characterized in that the first reagent solution and the second reagent solution are capable of forming a transparent or translucent mixed solution suitable for qPCR.

10. The solution set according to claim 2 , characterized in that the first reagent solution is selected from: polymerase solution, polymerase buffer; and the second reagent solution is selected from: primers, probe and sample.

11. The solution set according to claim 2 , wherein the absorbance peaks of any of said colorants do not overlap with the emission or excitation wavelength of the Alexa 350, FAM, JOE, VIC, HEX, Cy3, TAMRA, Cy3.5, Texas Red, ROX, Cy5, Cy5.5, Quasar705, or SYBR dye.

12. The solution set according to claim 2 , characterized in that the total absorbance of the mixed solution at the emission or excitation wavelength of the Alexa 350, FAM, JOE, VIC, HEX, Cy3, TAMRA, Cy3.5, Texas Red, ROX, Cy5, Cy5.5, Quasar705, or SYBR dye is less than 0.05, preferably less than 0.03, in particular less than 0.1 (1 mm light path).

13. Use of the method according to claim 1 , for preparing a reaction mixture for quantitative PCR.

14. Use of the solution set according to claim 1 , for preparing a reaction mixture for quantitative PCR.

Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Dec 8, 2021
From: TUUNANEN, JUKKA; KURKELA, JAAKKO; EKLIN, KATJA
To: FINNZYMES OY
Reel/Frame 058333/0228 →
BUSINESS TRANSFER AGREEMENT Recorded Dec 8, 2021
From: THERMO FISHER SCIENTIFIC OY
To: FERMENTAS UAB
Reel/Frame 058382/0858 →
MERGER Recorded Dec 8, 2021
From: FINNZYMES OY
To: THERMO FISHER SCIENTIFIC OY
Reel/Frame 058382/0954 →
CHANGE OF NAME Recorded Dec 8, 2021
From: FERMENTAS UAB
To: THERMO FISHER SCIENTIFIC BALTICS UAB
Reel/Frame 058382/0958 →
Priority Claims (1)
FI 20096013 · Oct 2, 2009 · national
Continuity (6)
Continuation 16815520 · Mar 11, 2020
Continuation 15989926 · May 25, 2018
Continuation 15210075 · Jul 14, 2016
Division 13078720 · Apr 1, 2011
Continuation In Part PCTFI2010050772 · Oct 4, 2010
Related Publication 20220275447A1 · Sep 1, 2022