METHOD AND SYSTEM FOR STORING INFORMATION BASED ON BIOMOLECULE

The present invention refers to 1) phase data binarized information by generating; 2) between the first and second probes binarized information by adding the step of inducing hybridization target strand; 3) hybridization by verifying the fluorescent signals; including a biomolecule based information to storage method are disclosed.

According to the method of the present invention biomolecule based information storage, the fluorescence signal at a simple mirror number switches short 25b is intended method may efficiently storing information as well as strand so as to control modification of the error, the large amount of information, can be efficiently storing data...copyright 2001.

More specifically said phase data binarized information to various disclosure information using fuzzy logic for generating a scheme commonly used but preferably using the ASCII code (ASCII). In the embodiment of the present invention whether one phase is encrypted as data in ASCII code KRIBB upon her.

Said 2) between the first and second probes binarized information by adding the step of inducing target strand for hybridization, hybridization probe assembly for between the first and second form can be prepared number is small. Said substrate is it was found the number can be used to secure the biomolecules comprising all material without substrate can be, for example nitrocellulose, nylon, beads, nanoparticles, cells, beads, one of glass may be the substrate disclosed.

Said probe is DNA, RNA, such as the purpose of the invention achieved single-stranded cDNA, hybridized form duplexes various biomolecule may be, corresponding to information bits (bit) different sequences can be characterized. Through each probe is complementary to the target sequence via addition specific hybridization is and can be, error includes a first current cut effective information encryption is the relationship.

Target strand is obtained in the present invention, between the first and second probes complementary hybridization of the head is can be DNA, RNA, such as the purpose of the invention achieved single-stranded cDNA, hybridized form duplexes can be a various biomolecules. When added to target strand, between the first and second probe hybridization is to take place, via the fluorescent hybridization to emit a signal to be coated. The amount of added target strand can be properly controlled according to the amount of data that is encrypted.

Said 3) hybridization step by verifying the fluorescent signals; on the substrate to prevent fluorescent signal in detecting the fluorescence signal is "ON" decrypting and, through encrypted information is recorded on the substrate can be confirmed. The fluorescence signal at a one-to-one match between the binarized information "1" which is "0" when "ON" of fluorescent signals is determined and represented "OFF" of fluorescent signals, "1" is determined as "0" when the opposite of fluorescent signals "OFF" of fluorescent signals is "ON" can be represented. Substrate which appears on the fluorescence signal binarized for decoding code which, in turn is in full text easily into a desired directional disclosed.

In addition the present invention refers to said biomolecule based information in storage method, after hybridization, substituted stranded on the substrate to a converted single-stranded adding hybridization strands; biomolecule based information storage further including number [...] substrate.

Said substituted stranded probe strand having a strand deteriorated compared to higher target 25b characterized and strand, error correction data via a re-write can be enabling. More specifically said substituted stranded deteriorated by the difference between the probe strand target molecule coupled to separate, isolated target strand and strand can be hybridized, competitive hybridization is measured for plural times. This process generates short called "toeholds" single domain, the processing advances through branched movement. Substituted strand length and/or sequence change controlled by the speed of the toehold by strand can be. In the present invention on a substrate comprising substituted stranded by converting single-stranded hybridization strands and the strand, through erroneous data or error for modifying error correction step to be characterized.

The present invention probe used, target strand, substituted stranded all single-stranded, such as stranded hybridization can be properly selected so that the biomolecule in constituting the power supply, preferably DNA, RNA, cDNA and consisting of at least one selected from the group consisting 1 be a.

In particular the present invention used in probe, target strand, substituted strand has an elongated sequences for many of the synthesis and the induced stress concentration through sequence analysis to resolve the door number, strand synthesis may occur in error is effectively lowered of oligonucleotide length ratio of 5 to 40 and can be, more preferably of at least 5 to 36 can be made. In addition said probe, target strand, 40 to 60% GC content is substituted strand, preferably 50% randomly to be synthesized.

The method of the present invention through two biomolecule based information storage method can exhibit fluorescence signal.

Fluorescent signal is labeled using hybridization occurs first target strand strands in fluorescent signal from a can, after stranded by substitution of a rewrite data, error correction added to labeled [khwen wife substituted strand can be made through signal to extinction.

The nucleotide sequence of the present invention target strand and fluorescent labeled, the labeled sequences can be substituted strand [khwen wife is composed.

Said fluorescence labelled sequences, the number is one [phul base five [ley new (fluorescein) but not limited to, chloro [phul base five [ley new tree oh it will keep (fluorescein chlorotriazinyl), with was and it pushed green (rhodamine green), red (rhodamine red) with was and it pushed, it was with a methyl and it pushed tetra (tetramethylrhodamine), FITC, [...] green (Oregon green), the rack which will know yarn fluoro (Alexa Fluor), FAM, JOE, ROX, HEX, various red (Texas Red), TET, TRITC, TAMRA, cyanine (Cyanine) based dye and the seed oh it is not a d car [lu time signal consisting of at least one selected from the group consisting 1 (thiadicarbocyanine) dye can be labeled sequences.

In addition said answer room [khwen wife labeled sequences (Dabcyl), TAMRA, Eclipse, DDQ, QSY, black parallel queen department (Blackberry Quencher), [...] queen department (Black Hole Quencher), Qxl, black (Iowa black) FQ iowa, iowa black RQ, IRDye QC-a 1 1 can be selected from the group consisting of at least one labeled sequence.

Said fluorescence labelled target strand and [khwen wife such as labeled substituted strand is applied, labeled sequences can be generated for the synthesis costs. The method of the present invention is a method for use in diameter than number based storage biomolecules, labeled sequences can for measuring fluorescent sequence and judged. For the method, addition of target sequence previously, simultaneously, fluorescent stage on a substrate after adding substrate. Fluorescent means substituted stranded, hybridization stranded, probes having high binding ability of the permutation is stranded - hybridization stranded - probe order characterized, in the presence of the probe and the probe - single-stranded target stranded hybridization strand bind to a double-stranded hybridization and organic, probe, hybridization stranded, substituted strand on a substrate in the presence of substituted stranded in conjunction with a stand-alone number fluorescence substrate.

Said fluorescent means the number is one but not limited to, SYBR Green provided 1, ethynyl galactooligosaccharides bromide, pico green, acridine orange, thiazole orange, YO provided PRO provided 1, at least one selected from the group consisting of chromia A3 1 be a yeast.

In addition the present invention refers to a stationary substrate probe, probe complementary target strands including biomolecules based information storage composition, kit and system are disclosed. To avoid complexity by redundant substrate on the same content with the specification description is given of a dispensed to each other.

The present invention according to system comprises a biometric molecules based data storage system is preferably microfluidic chips (microfulidics chip or lab provided on a-a-a chip) can be based data storage system. Said microfluidic chip-based information storage system includes implementing a lap fast, with high efficiency in the event that the probe of the present invention can analyze information related to fixed board and probe complementary target strands including a memory system.

Said microfluidic chip is it was found using the method publicly known number bath 1308.

Said composition, kit and system desired biomolecule hybridization data, encrypted effective via the strand to each other.

Composition of the present invention, kit and system complementary target 25b having high binding ability of the strands can be further substituted, through data in writing and error correction process from.

Said composition, kit and used in the system and the target strand fluorescence labelled strand, in this case [khwen home of wife's parents can be further labeled substituted strands.

In addition said composition, kit and system further comprises a stage can be fluorescent.

Instructions for use of the present invention optionally include a kit kit can be ascertain, probe are fixed substrate, the number 1 and number 2 compartment compartment separate probe complementary target strand preferably incorporated.

Hereinafter, the present invention in the embodiment and a process by the experiments detailed as follows. Stage, in the embodiment for the present invention is exemplified ephemeral embodiments and experiments, in the embodiment of the present invention and experiments and not limited to process by the contents.

In the embodiment. DNA - based storage device design

DNA based storage device for storing the desired information, data using fuzzy logic, fluorescent switch strategy based storage device DNA using front and rear. DNA encryption and decryption is based storage device comprises the step of, when a rewrite data to make data through a modifying step can be. In order to identify such concept, illustratively KRIBB Binary conversion table according as a desired full text is converted into ASCII code displayed as a transparent conductive layer, glass slide on a cryptographic process applied. After this, when the collecting fluorescent data Image, binary code for decrypting information conducting analysis. In addition KRIBT KRIBB full text in a desired full text conducting modifying data error - correction process.

DNA of fluorescent signals is based storage device which can exhibit data encrypted with the "ON" and "OFF", each "ON" and "OFF" performance "0" and "1" corresponding to each connection is implemented between the binary code with each other. The "0" functionalized single-stranded DNA probe by big fixed onto the surface state. The "1" state after hybridization or strands substituted one of double stranded DNA coupled and secured to the surface by big. "0" "1" "1" In the rewrite data "0" in DNA hybridization to DNA for performing conversion into a strand made by performing. The method also briefly a shown to 1.

As also shown in 1 information encrypted with the binary code "1" and then to desired DNA using fuzzy logic signal indicating signal representing "0" through "OFF" fluorescent "ON" and easily decryption with each other.

After oligonucleotide sequences used in example experiments as follows.

[Table 1]

[Khwen wife fluorescent stage and used substitution oligonucleotide sequences

[Table 2]

The full text of the aminated oligonucleotide library for encryption 0 bit 40 (36 oligonucleotide) sequences

[Table 3]

Full text 1 bit for encryption library oligonucleotides (26 oligonucleotide) sequence (* display from strand end up table 2 derived from a complementary oligonucleotide exhibits the same spot of 0 bit portion)

[Table 4]

A rewrite data for testing a second complementary oligonucleotide sequence permutations of oligonucleotides 36, 38 and 39 (26 nucleotide).

Experiment example 1. KRIBB encryption and decryption of data

1. 1 Immobilization and DNA hybridization probe for encryption

Table 1 total 40 shown functionalized amine group such as probe oligonucleotides modified and aldehyde coated glass slide that immobilize and 37 is in the him as time 1. Domain sequences Bioneer Corporation (Daejeon, South Korea) was synthesized in HPLC and positive number. Marienfeld Laboratory Glassware Company (Germany) to glass slide is purchased from the deflection, 3 a-aminopropyltriethoxysilane (APTES) was purchased from sigma is [...] solution and. Glycine seller deflection, saline-a sodium citrate (20X SSC solution) from the Ambion, 0.50 to seller, electrical resistance 18. 2 M Ω (Milipore, USA) obtained when the deionized water greater than Milli a-Q system, as well as after experiments.

Cleaning a surface immobilized buffer (Buffer 1: 1X SSC, 0. 1% SDS; Buffer 2:0. 1X SSC, 0. 05% SDS; Buffer 3:0. 1X SSC) is performed with respect to the transparent conductive layer using nitrogen air and dried. Additional time of 150 mm glycine blocking solution 1 was then surface. Finally said surface is performed carefully when the excess buffer solution and water and dried with respect to the nitrogen. For bit "0" spot, only single-stranded DNA probe applied only. Bit "1" for spot, complementary target strand (100 nm) applied to a DNA hybridization.

Spiral DNA (dsDNA) hybridize in double - buffer (10 mm Tris a-HCl, 50 mm NaCl, and 1 mm EDTA, pH 7. 8) Concentrations of probe DNA and target DNA in the same mixing of high pressure liquid coolant through his number. 5 Minutes to the mixture 90 °C have heating, cooling at room temperature when the 90 minutes, was stored at 4 °C for use later.

Spiral DNA (dsDNA) hybridize in double - buffer (10 mm Tris a-HCl, 50 mm NaCl, and 1 mm EDTA, pH 7. 8) Concentrations of probe DNA and target DNA in the same mixing of high pressure liquid coolant through his number. 5 Minutes to the mixture 90 °C have heating, cooling at room temperature when the 90 minutes, was stored at 4 °C for use later.

All the non - labeled DNA a 10 mm PBS buffer (pH 7. 4, 100 Mm NaCl) with respect to the primary dissolved. 0. 5 UM of aminated DNA solution (GA) activated glass slide on probe [...] 37 °C then applied during time in 1, 0. 01% (V/v) including 2X SSC was a solution of SDS (Saline provided Sodium Citrate) is performed. Capture DNA modified glass slide activated immediately soaked in 150 mm glycine solution in one time-gate 37 °C for blocking the binding site. 0 Continuously. 5 UM solution 45 °C target DNA in a DNA hybridization probe was two hours. DNA probes are not 0. 01% SDS (v/v) containing the hardening in 2X SSC and excess cleaning his number.

The scheme which results in an encrypted full text shown through the encrypted signal is also 2 "KRIBB" such as fluorescent DNA has been expected to be formulated. I.e., 2 as also shown in turn "1" character KRIBB into binary code that encrypted field signal "0" fluorescent "ON" fluorescent "OFF" field signal represented in detecting the fluorescence signal can be represented by DNA chip.

1. 2Through fluorescence measurement, decrypting encrypted data

All fluorescence measurement was performed using the quartz cuvette and fluorescence spectral with meter. Of 588 nm at room temperature using excitation wavelength, emission spectrum was collecting in the 800 nm to 595. The length of the excitation and emission slit [...] setting to 3 nm, 600 nm/minutes scanning rate electrodes respectively. In fluorescence intensity signal 607 nm fluorescent end ROX optimum one for a minor. Target and probe DNA composite hybrid buffer (10 mm Tris a-HCl, 50 mm NaCl, and 1 mm EDTA, pH 7. 8) Of a polymer of the same concentrations of target DNA and probe DNA before require - anneal process. Said mixture is 95 °C heating in, said equalizing temperatures after 15 minutes, 2 20 °C has been cooling slowly at a constant rate over time, was stored at 4 °C for use later. Said phosphor method such as result to identify via encrypted "KRIBB" 3 also shown. It is a woman wavelength of 488 nm fluorescent scanner Axon GenePix 4200A Image is obtained via a transparent conductive layer, in the emitting Sybr Green I 525 nm is known. Therefore, standard blue filter ((508 - 560 nm range) was selected.

As shown in fig. 3, causes the encrypted full text in "KRIBB" Image has been identified. 0 And 1 bit for each position clearly through glass slide on and OFF ON fluorescence signal has been identified. Simply, said bit is "0" only having only spot while the probe strand, said "1" bit is hybridization strength and, a mixture of "1" bit (final 20X concentration) target sequence Sybr Green I quadratures position are disclosed. I.e. the bit "1" exhibit fluorescence of Figure 3, the portion of the bit "0" exhibits fluorescence. Through, DNA based storage device storing and fluorescent signal can be effectively decrypt data has been confirmed.

Experiment example 2. A rewrite data using DNA strand

In information storage device, uncorrected data is repair or re-write the requirements can be very important since, in this regard in the embodiment when the write strategy such as data in establishing, one of the same two data for identifying first "KRIBB" wrong "KRIBT" the same method as example 1 character terms experiment was made via slide on an encrypted.

KRIBB KRIBT B T is stored in binary code is "1" to "0" according to error of protective switches 36 and 38 of which, is "0" "1" at position 39 to output an other. Such modifications in said two DNA hybridized DNA strands to form the substituted stranded by introducing form duplexes in separating strand can be composed of DNA stranded target DNA. In addition 39 position the oligonucleotide of complementary DNA hybridization can be accomplished by injecting inducing stranded oligonucleotides. The DNA strand and hybrid method using error correction and data rewrite to the concept shown also 4.

2. 1 Sybr Green I using data decrypting

Reduction of the construction cost in storage device using DNA is an important tube examination device are disclosed. The aminopeptidase to strand DNA synthesis and DNA produced in order to reduce the cost of the most adjacent cell is Sybr Green I can be using. For example O2 Sybr Green I used for detecting a nucleic acid, a single stranded DNA than a stronger binding affinity to bind with the unique double stranded DNA is known. Using Sybr Green I for, said probe DNA immobilized on the surface with respect to a regulation function distally amine aldehyde modified glass. Sybr Green I very low concentration (20x) reduction in costs by ray effectively can reduce...copyright 2001. Sybr Green I to decrypt information display, simply a fluorescent scanner (Axon GenePix 4200A scanner) is measured for plural times.

Sybr Green I and phosphor using method and apparatus for encoding data rewrite process 5 also to briefly shown. As shown in fig. 5, data "1" representing a binary code is binary fluorescent "ON", "OFF" is "0" by a goniophotometer. ". ". "". "0" Indicating "OFF" by forming a double-stranded DNA by hybridization of target DNA of superimposing more SybrGreen I high affinity to switch from "1" through connection that developing representative are disclosed. In addition DNA sequences hybridized double-stranded when substituted for a rewrite data added by single-stranded DNA probe signal "ON" is switched to the double stranded DNA target sequences and substituted by separating and Sybr Green I made to, have been converted to "0" through "OFF" signal encoding can be detoxification.

2. 2 Fluorescent stage and [khwen wife

Encryption and error (error-a correction) - modified first effort to test, for use strand DNA, DNA strand was testing. The same oligonucleotide strands and said table 1 used, substituted stranded concentration of 0, 1 uM, 0. 25 UM, 0. 5 UM, 1uM, 2 uM while increasing each 0. 5 UM probe strand when the tube containing the addition, of fluorescent signals change to 6 also shown.

As shown in fig. 6, replaces the fluorescent intensity reduced linear association between increasing the concentration strand has been confirmed. Substituted stranded probe strand half concentration of 0. 25 UM regulated tube is reduced as compared to the fluorescence intensity reaches a half. The concentration of the same level as in the concentration of probe strands stranded substituted 0. 5 UM increases to a plating solution is made, signal is reduced about 15 times has been confirmed. 1 UM substituted strands in a comparison sample tubes including tube 2 uM and very low signal about 100au and regulated 7500au 200au signal were identified. These results through injecting DNA single strands by substituting "1" substituted stranded hybridized DNA "ON" signal corresponding to "0" corresponding to "OFF" signal can be effectively modify has been confirmed. I.e., DNA strand by writing data through material has been confirmed.

2. 3 Sybr Green I determination of a data rewrite

(Rewriting) to determine whether a rewrite data by using Sybr Green I perform in order to identify, glass slide on reaction takes place in the strand have, cleaned and the remaining Sybr Green I via a number encoded target strand relates to substituted wherein the wetting ability. Sybr Green I has a very weak affinity because single stranded oligonucleotide probe. As shown by the target strand hybridization probe strand by indicating "ON" fluorescent reaction substituted strand by the addition of a probe strand from target strand, substituted strand and is protruded through the glass slide on separation of Sybr Green I in detecting the fluorescence signal is equal to disappear from the field. According to a method of such strand reaction results have shown to also 7.

As shown in fig. 7, is added to the hybridization occurs in lanes 2, 3 times the apparent target strand probe strand fluorescent signals is observed but, substituted strand is added by either a fluorescent signal is generated in lane 4, 5 substituted strand is not [...] "OFF (0)" state were identified. According to these results, using DNA strand Sybr Green I rewrite data via a reaction may be carried out effectively can be know.

2. 4 Write data retransmission through KRIBT error correction

DNA strand and method for retransmitting data writing can be carried out since the effective via the confirm, more specifically "KRIBT" fluorescent Image encoding encoding a rewrite data so that the desired accurate full text in "KRIBB" (error correction) by dielectrophoresis. "KRIBT" encoding "KRIBB" encryption code is to a "0" instead of "1" at position 37 and 36 is encrypted with the correct encryption system and, "0" instead of "1" to indicate position 39 is addressed by encrypted occurs. To correct for these errors, and single-stranded DNA strand when the complementary oligonucleotide implantation embodiment, shown to result also 8.

Also 8 as shown in the web page indicating the modified KRIBB KRIBT fluorescent Image data has been converted into a rewrite. 36 And 37 at "1" through "0" at position error signal is modified to indicate that his DNA strand. In addition 39 39 for DNA hybridization "0" position of error signal is new the [ley which will grow mote [tu single-stranded oligonucleotides complementary second position "1" has been modified by the implantation of oligonucleotides. These results indicate that 36, 38 and 39 to correspond with the positions the divert that combined second spot, as a result desired full text in "KIRBB" were identified. ASCii conversation table data on the full text of displayed as images obtained to can be decrypted. Conversion result in desired full text to "KRIBB" been decryption.

Experiment example 3. Based storage device also identifying specific DNA

Using Sybr Green I for determining specific hybridization efficiency, as shown in table 5 for testing the hybridization efficiency of the mismatched sequence the VIP, mismatch with respect to the number of increases from 0 to 6. According to observation of the efficiency of hybridization signal for increasing the number mismatch result 9 also shown.

[Table 5]

As shown in fig. 9, the displayed hybridization efficiency is reduced correspondingly increased number mismatch (T40 - T41 > T60>T0>T20>T10) is his. At the center of the sequence having only one mismatch is indicative of the intensity of weakest signal T10 while, from the centre of the strand of mismatch is installed at the 2 higher than T-a 20 T provided 10 is shown having the hybridization efficiency.