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Multipath pre-activating device, peptide nucleic acid preparation system, and pre-activating and synthetizing method

A pre-activation and multi-channel technology, which is applied in the field of biomacromolecule preparation, can solve the problems of uneven mixing of monomers and activators, affecting the activation effect, and reducing the coupling efficiency of monomers, so as to improve coupling efficiency and facilitate cleaning , to avoid the effect of operating errors

Active Publication Date: 2012-10-03
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although PNA is an analogue of DNA and polypeptides, since PNA is different from DNA or polypeptides in molecular structure and chemical properties, pre-activation of monomers is required during the synthesis of PNAs, and the existing DNA or polypeptide synthesizers do not have a single The body preactivation device mainly controls the activation by the residence of the monomer and the activator in the pipeline. The uneven mixing of the monomer and the activator will affect the activation effect and directly reduce the coupling efficiency of the monomer.

Method used

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  • Multipath pre-activating device, peptide nucleic acid preparation system, and pre-activating and synthetizing method
  • Multipath pre-activating device, peptide nucleic acid preparation system, and pre-activating and synthetizing method
  • Multipath pre-activating device, peptide nucleic acid preparation system, and pre-activating and synthetizing method

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Effect test

Embodiment 1

[0058] Such as figure 1 , figure 2 The present invention is composed of an inert gas source 2, a gas solenoid valve array 6, a multi-channel pre-activation device 13, a fluid solenoid valve array 25, and PLC4; the inert gas source 2 selects high-purity argon, helium or nitrogen, and The gas pipeline 3 is connected to the common air inlet 8 of the gas solenoid valve array to provide gas power for reagent delivery;

[0059] The gas solenoid valve array 6 is composed of a series of two-way or three-way solenoid valves. The control terminal 7 of each solenoid valve in the array is connected to the control output terminal 5 of the PLC in turn, and the common input of the array is The gas port 8 is connected to the inert gas source 2; the gas outlet 9 of one solenoid valve in the array is directly connected to the multi-channel pre-activation device 19, and the gas outlets 9 of the remaining solenoid valves are connected to the reagent bottle 11 through the control gas solenoid valve a...

Embodiment 2

[0064] Combine the working principle of the multi-channel pre-activation device 13 of the peptide nucleic acid preparation system of the present invention figure 2 Make the following explanation: In this example, the pre-activated peptide nucleic acid monomer T is taken as an example. Under the control of an external computer instruction, the PLC will follow the control output connection table in Appendix 1 to perform the following specific operations: (1) PLC Turn on the control output Y0, and turn on the air pressure balance switch 30 in the multi-channel pre-activation device. At this time, the multi-channel pre-activation device 13 is in a state that can receive reagents; (2) The PLC simultaneously opens the control output terminals Y5, Y6 and Y7 to open The solenoid valves in the gas solenoid valve array are respectively connected to the reagent bottle 11 containing peptide nucleic acid monomer T, alkali solution and activator; control the delivery of 0.1ml peptide nucleic ...

Embodiment 3

[0066] In this example, a 9-fluorenyloxycarbonyl (Fmoc) protected peptide nucleic acid monomer was used to synthesize a peptide nucleic acid single-stranded 5'-ATCG-3' as an example. Figure 4 The microsphere particles in middle a are used as the synthesis carrier, and a large number of microsphere particles are placed in the reaction tank. Input the peptide nucleic acid single-stranded sequence to be synthesized into the external computer program. After the external computer program completes the necessary initialization, the external computer will follow Figure 8 The program flow shown sends a series of instructions to the PLC, and the PLC performs the following specific tasks according to the received instructions: (The specific control objects of the PLC can be seen in Table 1)

[0067] (1) Counter i=1, where i corresponds to the i-th position of the peptide nucleic acid sequence, for example, the peptide nucleic acid monomer corresponding to i=2 is T;

[0068] (2) Deprotectio...

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Abstract

The invention discloses a multipath pre-activating device, a peptide nucleic acid preparation system, and a pre-activating and synthetizing method. The peptide nucleic acid preparation system comprises the multipath pre-activating device, and further comprises an inert gas source, a gas electromagnetic valve array, a fluid electromagnetic valve array, a PLC (programmable logic controller), a computer, a reaction tank and a plurality of reagent bottles. The synthetizing method comprises a method for synthetizing peptide nucleic acid single-chain and a method for synthetizing peptide nucleic acid arrays. The multipath pre-activating device can effectively perform quantifying, and periodically mixing and activating to monomer reagents, and increase coupling reaction efficiency; at the same time, the multipath pre-activating device can coordinate with the electromagnetic valve arrays to realize multi-flow-path liquid transferring control, and can be cleaned easily. The device, system and method provided by the invention can meet demand on synthesis of peptide nucleic acids, and can be used for synthesis of peptide nucleic acid single-chain and peptide nucleic acid arrays.

Description

Technical field [0001] The invention belongs to the technical field of biological macromolecule preparation, and particularly relates to a multi-channel pre-activation device, a peptide nucleic acid preparation system and a pre-activation and synthesis method. Background technique [0002] Peptide nucleic acid (PNA) is an analog of DNA. The backbone of PNA is formed by linking N-(2-aminoethyl)glycine and nucleic acid bases through methylene carbonyl groups. PNA can specifically hybridize with DNA or RNA to form a stable complex. At the same time, PNA as a hybridization probe greatly improves the efficiency and sensitivity of genetic testing and medical diagnosis. [0003] At present, there is still no PNA synthesis equipment in the domestic and foreign markets. The synthesis of PNA is similar to the synthesis of DNA and peptides. It requires several cycles of deprotection, coupling, and capping reactions. Therefore, the existing PNA preparation is mainly synthesized by DNA or pep...

Claims

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Application Information

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IPC IPC(8): C07K1/00C07K14/00C40B50/06
Inventor 刘正春邬燕琪王宏银杨飞鹏
Owner CENT SOUTH UNIV
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