JP2003083967A - Biologically related substance fixed gel and biologically related substance fixed gel chip having the gel held thereto - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a gel composition accelerating the diffusion speed of a biologically related substance being a specimen without lowering the concentration of a gel and easy to form a hybrid, a biologically related substance fixed gel based on the gel composition and a biologically related substance fixed gel chip having the gel held thereto. SOLUTION: The biologically related substance fixed gel is prepared by fixing the biologically related substance within an aqueous gel containing 3-20 wt.% of an acrylamide type polymer and 0.1-10 wt.% of a polymeric polyhydric alcohol. The biologically related substance fixed gel chip wherein the gel is held to a plurality of sections is also disclosed.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a bio-related substance-immobilized gel containing a high molecular weight polyhydric alcohol and a bio-related substance-immobilized gel chip using the same. The chip is used for gene expression analysis and the like.

[0002]

2. Description of the Related Art In recent years, genome projects in various organisms have been under way, and many genes including human genes and their nucleotide sequences are rapidly being clarified. The function of the gene whose sequence has been elucidated can be examined by various methods. As one of the effective methods, gene expression analysis using the revealed nucleotide sequence information is known. For example, a method utilizing various nucleic acid: nucleic acid hybridization reactions and various PCR reactions represented by Northern hybridization has been developed, and the relationship between various genes and their biological function expression is investigated by the method. You can Although there is a limit to the number of genes that can be applied by these methods, the comprehensive number of genes at the single-individual level, which is being clarified through today's genome projects,
DNA microarray method (DNA chip method) that enables batch expression analysis of multiple genes for systematic analysis
A new analytical method, or methodology, has been developed.

As a DNA microarray, DN is formed on a large number of compartmentalized cells by applying lithography technology.
A synthesized (Science 251 767
-773), a partition is formed on the base by grooves or holes, and a probe is fixed to the inner wall of the partition (Japanese Patent Laid-Open No. 11-10).
8928, JP-A-2000-78998), in order to increase the amount of probe immobilized on the chip, a microarray in which the probe is immobilized on a gel such as acrylamide (US
P 5,770,721, Japanese Patent Laid-Open No. 2000-60554.
No.) and many other shapes are known. Some of the present inventors previously developed a novel microarray and a method for manufacturing the same (Japanese Patent Laid-Open Nos. 2000-270878 and 2000-
270879). According to the invention, a thin piece is obtained by producing a nucleic acid-immobilized gel-holding fiber array holding a nucleic acid-immobilized gel and cutting the array in a direction intersecting the fiber axis of the array. This thin piece is used as an immobilized nucleic acid two-dimensional high-density array, that is, a DNA microarray.

[0004]

A microarray in which probes are immobilized on a gel such as acrylamide has a three-dimensional immobilization field as compared with an array in which the probes are immobilized on a non-porous substrate, and therefore has a large area in a small area. Can hold the probe. However, the bio-related substance that acts as a sample to act on the microarray needs to diffuse into the gel and form a hybrid inside the gel. Therefore, the detection rate of the sample is diffusion-controlled, and the diffusion rate is slower for higher molecular weight samples. Therefore, there is a problem in that it takes time to detect.

Therefore, in the case of a bio-related substance-immobilized chip using a gel, it has been required to increase the diffusion rate of a higher molecular weight sample into the gel, accelerate the hybrid formation rate, and increase the detection rate. . As a means for solving this problem, a method of lowering the gel concentration can be considered, but if the gel concentration is too low, the gel becomes soft, and if the acrylamide concentration is 4% or less, it is difficult to form a gel. On the other hand, in the field of electrophoresis gel, biological substances such as proteins and D
To increase the moving speed of NA, etc., Japanese Patent Laid-Open No. 8-15222
Although monomers other than acrylamide have been proposed, such as No. 2, a medium comparable to acrylamide gel has not been developed because of its gel strength, polymerization rate, and the like.

According to the present invention, a gel composition in which a bio-related substance as a sample accelerates the diffusion rate and easily forms a hybrid without lowering the gel concentration, and a bio-related substance-immobilized gel based on this composition and the gel are retained by the gel. An object of the present invention is to provide a biochip-immobilized gel chip.

[0007]

Means for Solving the Problems As a result of intensive studies for solving the above-mentioned problems, the present inventors have found that a biological substance in which a biological substance is held in a gel containing a specific amount of an acrylamide polymer and a high molecular weight polyhydric alcohol. Related substance-immobilized gel
The present inventors have found that the diffusion rate of a biologically-relevant substance as a sample can be markedly increased as compared with a conventional gel, and the hybridization rate in the gel can be improved, and the present invention has been completed.

That is, the present invention uses 3 to 20% by weight of an acrylamide polymer and 0.1 of a high molecular weight polyhydric alcohol.
A bio-related substance-immobilized gel in which a bio-related substance is immobilized in an aqueous gel containing 10% by weight. Examples of the high molecular weight polyhydric alcohol include at least one selected from the group consisting of compounds represented by the following general formulas (1) to (3). Formula _{(1) OH- [CH 2 CH} 2 O-] nH ( wherein, n is an integer of 10 or more) Formula _{(2) OH- [CH 2 (} CH) CH 3 O-] nH ( Formula In the formula, n is an integer of 10 or more. General formula (3) ROCH ₂ CHORCH ₂ OR (In the formula, R represents [-CH ₂ CHCH ₃ O] nH, and n is an integer of 5 or more) The molecular weight of the high molecular weight polyhydric alcohol is usually 1,000 to
It is in the range of 1,000,000. In addition, examples of the biological substance include nucleic acids.

Further, the present invention is a bio-related substance-immobilized gel chip in which the bio-related substance-immobilized gel is held in a plurality of compartments. The area of each of the plurality of sections is usually 10 ⁻⁶ m ² or less. These compartments are, for example,
Hollow fibers can be used as the material for forming the through holes, which are formed by grooves or through holes.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION The gel for immobilizing a biological substance of the present invention comprises 3 to 20% by weight, preferably 4 to 15% by weight, of an acrylamide polymer and 0.1% of a polyhydric alcohol.
An aqueous gel containing 10 to 10% by weight, preferably 1 to 10% by weight.

The acrylamide polymer is a copolymer composed of at least one monomer of (meth) acrylamide and N-substituted (meth) acrylamide and a crosslinking agent. Examples of the monomer include acrylamide, methacrylamide, N.I. N-dimethylacrylamide, N-
Examples include isopropyl acrylamide and N, N-diethyl acrylamide. The crosslinking agent is not particularly limited as long as it is copolymerizable with these monomers. For example, methylene bis acrylamide, bis acrylamide methyl ether, (poly) ethylene glycol di (meth) acrylate and the like which are generally used in electrophoresis gel and the like can be mentioned. The amount of the crosslinking agent used is 1 to 10 with respect to the monomer.
Weight percent is preferred.

The high molecular weight polyhydric alcohol is a compound having a plurality of OH groups represented by the above general formulas (1) to (3). Examples thereof include polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxymethyl cellulose and the like. The molecular weight of these high molecular weight polyhydric alcohols is 1,000 to 1,000,000, preferably 5,000 to 100,000.

Examples of the bio-related substance to be immobilized on the gel include deoxyribonucleic acid (DNA) and ribonucleic acid (RNA). The bio-related substance used in the present invention may be either a commercial product or a bio-related substance obtained from living cells or the like.

For example, when a nucleic acid is used as the biological substance, a known method for preparing DNA or RNA from living cells, for example, for the extraction of DNA, the method of Blin et al. (Nucleic Acids Res. 3.2) is used.
303 (1976)) and for the extraction of RNA, the method of Favaroro et al. (Method).
s. Enzymol. 65.718 (1980)) and the like. Furthermore, linear or circular plasmid DNA or chromosomal DNA is used. These DNAs are DNAs cut with restriction enzymes or chemically.
It is also possible to use fragments, DNA synthesized by an enzyme or the like in a test tube, or chemically synthesized oligonucleotides.

In the present invention, the bio-related substance may be directly immobilized on the gel, or a derivative obtained by chemically modifying the bio-related substance, or a bio-related substance modified as necessary may be immobilized. Good. For immobilizing the bio-related substance on the gel, a method of physically encapsulating in the gel or a direct binding to the gel component may be used. The carrier may be bound to the carrier by covalent bond or non-covalent bond, and the carrier may be immobilized on the gel. For example, it is possible to introduce a vinyl group into the terminal group of a biological substance and copolymerize it with a gel component such as acrylamide (WO98 / 39351).

The bio-related substance-immobilized gel thus prepared is, for example, USP 5,770,721, JP 2000-60554 A, JP 2000-27087 A.
No. 8, DNA described in JP 2000-270879 A, etc.
It can be used as a gel for holding microarray probes.

For example, in a gel chip on which a biological substance is immobilized, a substrate having a smooth surface, a substrate having a plurality of grooves, a substrate having a plurality of through holes, etc. can be used as a substrate for holding the gel. For example, when a substrate having a plurality of grooves or through holes is used, the bio-related substance-immobilized gel monomer solution before or immediately after the initiation of polymerization is added to the section formed by the grooves or the through holes to gel within the section. By carrying out the above, a gel chip on which a biological substance is immobilized can be prepared. An example of the substrate having a plurality of through holes is a hollow fiber array body sliced in a direction intersecting the fiber axis (Japanese Patent Application No. 11-93044). The types of bio-related substances retained in each compartment may be different or may be grouped. Further, as a positioning index, for example, a gel in which a dye is retained can be retained in the compartment instead of the biological substance. Also,
The area of each of the plurality of sections is usually 10 ⁻⁶ m ² or less. The lower limit is not particularly limited as long as it is possible to detect a biological substance.

The bio-related substance-immobilized gel obtained in the present invention and a chip in which the gel is divided into a large number of compartments react with a specimen using the immobilized bio-related substance as a probe, just like the gel chip shown in the prior art. By carrying out the hybridization, it can be used for detection of a biological substance having a specific base sequence in a sample. In the broad sense, the term “probe” as used in the present invention refers to a bio-related substance capable of specifically binding to a protein, a low molecular weight compound, or the like present in a sample. That is, the bio-related substance-immobilized gel of the present invention is reacted with a sample to perform hybridization, form a hybrid with a bio-related substance present in a sample complementary to the probe, and detect the hybrid It is possible to detect a biological substance in the sample.

For detecting the hybrid, a known means capable of specifically recognizing the hybrid can be used. For example, it is possible to introduce a fluorescent substance into a biological substance as a sample and detect the hybridization reaction amount between the sample and the probe as the fluorescence intensity introduced into the sample.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, the technical scope of the present invention is not limited by these examples.

Reference Example 1 (a) Preparation of Vinyl Group-Introduced Oligonucleotide Probes The following oligonucleotides (Probe A and Probe B) were synthesized. Probe A: cgcgttacct tccgcgagat cagtatccac cga (SEQ ID NO: 1) The oligonucleotide was synthesized by PE Biosystems' automatic synthesizer DNA / RNA synthesizer.
(Model 394), and in the final step of DNA synthesis, NH ₂ (CH ₂ ) ₆ − was added to the 5′-end of each oligonucleotide using Aminolink II (trade name) (Applied Biosystems). An introduced and aminated probe was prepared. These were used after being deprotected and purified by a general method. Obtained probe A (1
000 nmol / ml) 10 μl, 1000 mM sodium carbonate aqueous solution 5 μl, and methacrylic anhydride 5 μl dissolved in DMSO at a concentration of 20 μmol / ml were mixed and reacted at room temperature for 2 hours to synthesize a vinyl group-introduced oligonucleotide probe. did.

(B) PCR Reaction of Oligonucleotide Introducing Terminal Vinyl Group (b-1) Preparation of Template (Chromosome) Rhodococcus rhodochrous J-1 strain was fed to a nutrient medium (glucose 15 g, yeast extract 1 g, sodium glutamate 10 g, KH ₂ PO). ₄ 0.5g, K ₂ HPO ₄
0.5 g, MgSO4.7H2O 0.5 g / L, pH
7.2) The cells were cultured in 100 ml at 30 ° C. for 3 days to collect the cells.
A chromosome was prepared from this cell and used as a template for PCR.
In addition, Rhodococcus rhodochrous J-1 strain is FERM
Deposited as BP-1478 at the Institute of Biotechnology, Institute of Industrial Science and Technology (1-3, Higashi 1-3, Tsukuba, Ibaraki).

(B-2) PCR The terminal vinylated oligonucleotide (SEQ ID NO: 1) prepared in Reference Example was diluted to 50 μM with sterile water, and the oligonucleotide (SEQ ID NO: 2) was diluted to 5 μM with sterile water.
PCR reaction was performed using the template prepared in (1). Oligonucleotides were commissioned to Amersham Pharmacia for synthesis. PCR was performed in accordance with the specifications of Ex-Taq (Takara Shuzo), TaKaRa PCR Thermal Cycle.
er PERSONAL. 100 reactions
μl, temperature conditions are 93 ° C. for 30 seconds, 65 ° C. for 30 seconds
35 cycles of 1 second at 72 ° C. for 1 second. By this reaction, the 623 base (SEQ ID NO: 3) 5'-terminal vinylated nucleic acid (probe C) was amplified.

(C) Preparation of sample nucleic acid (sample) (c-1) Synthesis of oligonucleotides The following oligonucleotides (probe B, probe D) were synthesized. Probe A: cgcgttacct tccgcgagatc agtatccac cga (SEQ ID NO: 1) Probe B: cgcgacaatg gcgagcttgt attcaccgag (SEQ ID NO: 2) Oligonucleotide synthesis was performed in the same manner as in (a), and Cy5 was added to the 5'end of the oligonucleotide (probe B). A model of the introduced fluorescence-labeled sample nucleic acid was prepared. These were used after being deprotected and purified by a general method.

(C-2) PCR The 5'-end Cy5-labeled oligonucleotide (SEQ ID NO: 2) described above was diluted to 50 μM with sterile water, and the oligonucleotide (SEQ ID NO: 5) was diluted to 5 μM with sterile water, 1)
And the template (SEQ ID NO: 2,
PCR reaction was carried out using 5). PCR reaction is Ex
-TaKaRa P according to the specifications of Taq (Takara Shuzo)
CR Thermal Cycler PERSONA
Performed using L. The reaction was carried out at 100 μl, and the oligonucleotide (SEQ ID NO: 2) temperature condition was 93 ° C. for 30 seconds,
35 cycles of 65 ° C. for 30 seconds and 72 ° C. for 1 minute were performed.
By this reaction, part of 623b (SEQ ID NO: 3) (22
8 to 623), a 5'-terminal Cy5-labeled single-stranded nucleic acid (395 base), which is a complementary sequence, was amplified.

<Example 1> (1) Preparation of model gel with bio-related substances immobilized (1-1) Preparation of monomer solution A monomer solution having the composition shown in Table 1 was prepared.

[Table 1]

(1-2) Preparation of gel Immediately after preparation of the monomer solutions shown in Table 1, the length was 2 cm.
10 μl was added to a 1 mmφ hematocrit tube cut into 1 and left at room temperature for 1 hour to prepare an acrylamide gel in which the terminal vinyl group nucleic acid obtained in Reference Example (a) was immobilized.

(2) Hybridization The hybridization solution shown in Table 2 was mixed with the sample nucleic acid (specimen) solution as shown in Table 3 and the model gel prepared in (1-2) was placed in a 1500 μl capacity microtube. It was added to the sample solution. Hybridization was carried out by leaving it in an incubator adjusted to 60 ° C. for about 20 hours.

[Table 2]

[Table 3]

(3) After washing and hybridization, the hematocrit tube was taken out from the microtube, transferred to 50 ml of 0.1 × SSC, 0.1% SDS solution which had been kept warm, and shaken at 45 ° C. at 20 ° C. Washing for 3 minutes was performed 3 times.

(4) The gel after completion of the detection washing was observed with a fluorescence microscope (Olympus fluorescence microscope BX60) using a cy5 filter,
The fluorescence in the depth direction of the gel (that is, the longitudinal direction of the hematocrit tube) was observed, and the cooled CCD camera (C by Olympus Co., Ltd.
Images were acquired by OOLSnap). With respect to the obtained image, a line profile of fluorescence intensity in the longitudinal direction of the hematocrit tube was obtained by image analysis software Image-Pro P.
It was measured with lus (manufactured by Planetron), and the integrated value of the fluorescence intensity from the gel surface was measured. Comparative Example 1 Gel preparation, hybridization, and detection were performed in the same manner as in Example 1 except that polyethylene glycol 20000 was omitted.
The results are shown in Table 4.

[0031]

[Table 4] *) The higher the integrated value of the fluorescence intensity, the more the amount of hybrids formed.

Reference Example 2 (a) Preparation of vinyl group-introduced oligonucleotide probe, (b) PC of terminal vinyl group-introduced oligonucleotide
The R reaction was performed in the same manner as in Reference Example 1. (C) Preparation of sample nucleic acid (sample) (c-1) Synthesis of oligonucleotide The oligonucleotides (probe B, probe D) shown below were synthesized. Probe A: cgcgttacct tccgcgagat cagtatccac cga (SEQ ID NO: 1) Probe C: gactcggggc tgatcagcga agatgagatc (SEQ ID NO: 4) Oligonucleotides are synthesized by PE Biosystems automated synthesizer DNA / RNA synthesizer.
(Model 394). Regarding the oligonucleotide (probe C), a model of a fluorescence-labeled sample nucleic acid in which Cy5 was introduced at the 5'end was prepared. They are,
It was used after being deprotected and purified by a general method. (C-2) PCR The 5 ′ unterminated Cy5-labeled oligonucleotide (SEQ ID NO: 2) described in C-1 was diluted to 50 μM with sterile water, and the oligonucleotide (SEQ ID NO: 1) was diluted to 5 μM with sterile water,
A PCR reaction was performed using the template prepared in (1) and the oligonucleotides (SEQ ID NOs: 1 and 2). The PCR reaction is
According to the specifications of Ex-Taq (Takara Shuzo), TaKaRa
PCR Thermal Cycler PERSO
This was done using NAL. The reaction was carried out in 100 μl, and the oligonucleotide (SEQ ID NO: 2) temperature condition was 93 ° C. 30
Seconds, 65 ° C. for 30 seconds, 72 ° C. for 1 minute for 35 cycles. By this reaction, the 5'terminal Cy which is a sequence complementary to a part (420 to 623) of 623b (SEQ ID NO: 3).
5-labeled single-stranded nucleic acid (203 base) was amplified.

<Example 2> (1) Preparation of fiber array body ^Two fiber guide plates each having a total of 25 holes arranged vertically and horizontally on a square of 1 cm ² were used, and two fiber guide plates were regularly arranged. A hollow fiber array was obtained by passing a total of 25 nylon hollow fibers (outer diameter of about 300 μm, length of about 50 cm) in five vertical rows and five horizontal rows. A hollow fiber array having a portion which is not fixed by resin at both ends by pouring a polyurethane resin (Nippon Polyurethane Industry Co., Ltd., Corona 4403, Nipporan 4223) between the two fiber guide plates at a distance of 20 cm and curing it. Got

(2) Introduction and Immobilization of Biopolymer into Hollow Fiber Arrangement Monomer solutions 3 and 4 having the following composition, including those synthesized in Reference Example 2, were prepared.

[Table 5]

The monomer solution 3 or the monomer solution 4 is
After injecting into the hollow part of the hollow fiber of the hollow fiber array, it was transferred to a closed glass container whose inside was saturated with water vapor, and left at 80 ° C. for 4 hours to carry out a polymerization reaction. As a result, a hollow fiber array having a gel having terminal vinylized nucleic acid immobilized therein was obtained.

(3) Preparation of biopolymer array thin piece:
The terminal vinylated nucleic acid-immobilized hollow fiber array obtained in (2) is cut out in a direction perpendicular to each fiber axis to a thickness of about 500 μm using a microtome, whereby the terminal vinylated nucleic acid becomes regular. A biopolymer array thin piece was obtained.

(4) Hybridization Table 5 is attached to the hybrid pack (manufactured by NIPPON GENETICS CO, LTD).
100 μl of the hybridization solution shown in 1) was added, and the biopolymer array thin piece prepared in (3) was added to the hybridization solution and heat-sealed. 65
Hybridization was carried out by leaving it in a shaker adjusted to 0 ° C. for about 20 hours.

[0038]

[Table 6] (5) Washing After completion of the hybridization, the biopolymer array thin piece was taken out from the hybrid pack and the following washing operation was performed. 2 × SSC, 0.1% SDS room temperature 20 minutes 0.2 × SSC, 0.1% SDS 55 ° C. 20 minutes 0.2 × SSC, 0.1% SDS 55 ° C. 20 minutes 0.02 × SSC Room temperature 5 minutes 0.02 × SSC Room temperature 5 minutes

(6) The thin piece after completion of the detection and washing was observed with a fluorescence microscope (Olympus fluorescence microscope BX60) using a cy5 filter,
Cooled CCD camera (Olympus COOLSnap)
I got the image at. The exposure time was 15 seconds. For the obtained image, the average value of the fluorescence intensity in the spot where the terminal vinylated nucleic acid was immobilized was calculated. The image analysis was performed using image analysis software Image-ProPlus (manufactured by Planetron). The results are shown in Table 6.

<Comparative Example 2> In the monomer solution composition of Example 2, except that polyethylene glycol 20000 was omitted, the biopolymer was introduced into and immobilized on the hollow fiber array in the same manner as described above. Was prepared, hybridized, washed, and detected. The detection results of Example 2 and Comparative Example 2 are shown in Table 6.

[0041]

[Table 7]

[0042]

EFFECT OF THE INVENTION The gel composition of the present invention has a higher hybrid formation amount between the sample and the probe as compared with the gel to which the high molecular weight polyhydric alcohol is not added. That is, it is possible to improve the speed of forming a hybrid.

[0043]

[Sequence list] <110> Mitsubishi Rayon Co., Ltd. <120> Bio-related material-fixed gel and bio-related material-fixed gel        chip holding said gel <130> <210> 1 <211> 33 <212> DNA <213> Artificial Sequence <223> Synthetic DNA <400> 1      cgcgttacct tccgcgagat cagtatccac cga <210> 2 <211> 30 <212> DNA <213> Artificial Sequence <223> Synthetic DNA <400> 2       cgcgacaatg gcgagcttgt attcaccgag <210> 3 <211> 623 <212> DNA <213> Rhodococcus rhodochrous <400> 3 gcgataggtg gaggtcaggg tttgggacag cagctccgaa atccgctaca tcgtcatccc 60 ggaacggccg gccggcaccg acggttggtc cgaggaggag ctgacgaagc tggtgagccg 120 ggactcgatg atcggtgtca gtaatgcgct cacaccgcag gaagtgatcg tatgagtgaa 180 gacacactca ctgatcggct cccggcgact gggaccgccg caccgccccg cgacaatggc 240 gagcttgtat tcaccgagcc ttgggaagca acggcattcg gggtcgccat cgcgctttcg 300 gatcagaagt cgtacgaatg ggagttcttc cgacagcgtc tcattcactc catcgctgag 360 gccaacggtt gcgaggcata ctacgagagc tggacaaagg cgctcgaggc cagcgtggtc 420 gactcggggc tgatcagcga agatgagatc cgcgagcgca tggaatcgat ggccatcatc 480 gactgacatc ccctgtgtct ccatctagca gcagtgcggg cgtaccccga cggtgctgag 540 ccgacggggt acgcccgcac ttcatcaatg acggtggttc ctaatttggc tcggtggata 600 ctgatctcgc ggaaggtaac gcg 623 <210> 4 <211> 30 <212> DNA <214> Artificial Sequence <223> Synthetic DNA <400> 4       gactcggggc tgatcagcga agatgagatc

[0044]

[Sequence list free text]

SEQ ID NO: 1 synthetic DNA SEQ ID NO: 2: Synthetic DNA SEQ ID NO: 4: Synthetic DNA

Claims (8)

[Claims] 1. A bio-related substance-immobilized gel in which a bio-related substance is immobilized in an aqueous gel containing 3 to 20% by weight of an acrylamide polymer and 0.1 to 10% by weight of a high molecular weight polyhydric alcohol. 2. The high molecular weight polyhydric alcohol is represented by the general formula (1) to
The bio-related substance-immobilized gel according to claim 1, which is at least one selected from the group consisting of the compounds represented by (3). Formula _{(1) OH- [CH 2 CH} 2 O-] nH ( wherein, n is an integer of 10 or more) Formula _{(2) OH- [CH 2 (} CH) CH 3 O-] nH ( Formula In the formula, n is an integer of 10 or more. General formula (3) ROCH ₂ CHORCH ₂ OR (In the formula, R represents [-CH ₂ CHCH ₃ O] nH, and n is an integer of 5 or more) 3. The high molecular weight polyhydric alcohol has a molecular weight of 1,0.
The bio-related substance-immobilized gel according to claim 2, which is from 0 to 1,000,000. 4. The biological-related substance is a nucleic acid.
9. The bio-related substance-immobilized gel according to any one of 1. 5. A bio-related substance-immobilized gel chip in which the bio-related substance-immobilized gel according to claim 1 is held in a plurality of compartments. 6. The area of each of the plurality of compartments is 10 ⁻⁶ m
^The biochip-related substance-immobilized gel chip according to claim 5, which is ² or less. 7. The biochip-immobilized gel chip according to claim 5, wherein the partition is formed by a groove or a through hole. 8. The bio-related substance-immobilized gel chip according to claim 7, wherein the through holes are formed of hollow fibers.