Office Action 1: Notice of Allowance (2025-02-04)

CLAIMS:
CLAIMS
1\. A method of gene expression analysis, comprising: a) contacting a first RNA analyte and a second RNA analyte in a cell or tissue sample with a first probe and a second probe, respectively, to generate a first amplicon and a second amplicon of a library of amplicons, wherein the first probe is a first padlock probe comprising a 3 ’hybridization region and a 5’ hybridization region complementary to the first RNA analyte in the arms of the first padlock probe and the second probe is a second padlock probe comprising a 3 ’hybridization region and a 5’ hybridization region complementary to the second RNA analyte in the arms of the second padlock probe, wherein the first amplicon comprises i) a first sequencing priming site and ii) a first identifier sequence, and the second amplicon comprising i) a second sequencing priming site different from the first sequencing priming site and ii) a second identifier sequence, wherein the first and second identifier sequences comprise a sequence derived from the first RNA analyte and the second RNA analyte, respectively; b) using a first sequencing primer to sequence the first identifier sequence of the first amplicon, c) removing, cleaving, or blocking the extension product of the first sequencing primer; and d) using a second sequencing primer different from the first sequencing primer to sequence the second identifier sequence of the second amplicon, thereby using the sequenced first and second identifier sequences to identify the first RNA analyte and the second RNA analyte.
2\. The method of claim 1, further comprising ligating a 3’ end and a 5’ end of the first probe using the first RNA analyte as template to form a first circularized template and ligating a 3’ end and a 5’ end of the second probe using the second RNA analyte as template to form a second circularized template.
3\. The method of claim 2, wherein the first circularized template and the second circularized template is used to perform rolling circular amplification to generate the first amplicon and the second amplicon, respectively.
4\. The method of claim 1, wherein the sequencing comprises performing sequencing-by-synthesis (SBS), sequencing-by-binding (SBB), or sequencing-by-avidity (SBA) of the first and second identifier sequences.
5\. The method of claim 1, wherein the sequencing comprises using a polymerase to incorporate a plurality of cognate nucleotides into the first sequencing primer or an extension product thereof and wherein the sequencing comprises using a polymerase to incorporate an additional plurality of cognate nucleotides into the second sequencing primer or an extension product thereof.
6\. The method of claim 5, wherein the plurality of cognate nucleotides and additional plurality of cognate nucleotides comprise a plurality of reversible terminator nucleotides coupled to a label via a linker.
7\. The method of claim 1, wherein b) and d) each comprises contacting the cell or tissue sample with a nucleotide mix comprising a fluorescently labeled nucleotide and a nucleotide that is not fluorescently labeled.
8\. The method of claim 1, wherein b) and d) each comprises contacting the cell or tissue sample with one of the following nucleotide mixes: nucleotide mix 1 in which nucleotides comprising G are not detectably labeled, whereas nucleotides comprising A, C, or T are detectably labeled; nucleotide mix 2 in which nucleotides comprising T are not detectably labeled, whereas nucleotides comprising A, C, or G are detectably labeled; nucleotide mix 3 in which nucleotides comprising C are not detectably labeled, whereas nucleotides comprising A, G, or T are detectably labeled; or nucleotide mix 4 in which nucleotides comprising A are not detectably labeled, whereas nucleotides comprising G, C, or T are detectably labeled.
9\. The method of claim 8, wherein independent of one another, in each nucleotide mix, the detectably labeled nucleotides comprise: i) fluorescent labels of three different colors, one for each of the three bases in the detectably labeled nucleotides; ii) fluorescent labels of two different colors, one each for two of the three bases in the detectably labeled nucleotides, wherein nucleotides comprising the remaining base are labeled with both colors; or iii) fluorescent labels of the same color, wherein fluorescent labels on nucleotides comprising one of the three bases in the detectably labeled nucleotides are configured to be cleaved, and nucleotides comprising another one of the three bases are configured to be labeled with the fluorescent label.
10\. The method of claim 1, wherein the first sequencing priming site is assigned to the first RNA analyte and the second sequencing priming site is assigned to the second RNA analyte based on a decision rule designed to minimize a maximum predicted density of signals detected in each sequencing cycle of nucleotide incorporation of the sequencing in the cell or tissue sample.
11\. The method of claim 10, wherein the decision rule uses expression data for the first RNA analyte and the second RNA analyte.
12\. The method of claim 10, wherein the assigning comprises assignment based on expression data for the first RNA analyte and the second RNA analyte in clustered cell types.
13\. The method of claim 12, wherein the expression data for the first RNA analyte and the second RNA analyte comprises spatial gene expression data, spatial protein expression data, single cell gene expression data, single cell protein expression data, or any combination thereof.
14\. The method of claim 1, wherein the cell or tissue sample comprises a first plurality of amplicons that share a common first sequencing priming site and a second plurality of amplicons that share a common second sequencing priming site.
15\. The method of claim 14, wherein: the first plurality of amplicons comprises identifier sequences from two or more different RNA analytes; and/or the second plurality of amplicons comprises identifier sequences from two or more different RNA analytes.
16\. The method of claim 1, wherein b) and d) each comprises contacting the cell or tissue sample with a first nucleotide mix in a first sequencing cycle and contacting the cell or tissue sample with a second nucleotide mix in a second sequencing cycle, wherein the first nucleotide mix and the second nucleotide mix each comprises nucleotides of four different bases and nucleotides of three different bases are detectably labeled and nucleotides of the other base are not detectably labeled.
17\. The method of claim 16, wherein the base of the nucleotides that are not fluorescently labeled in the second nucleotide mix is different from the base of the nucleotides that are not fluorescently labeled in the first nucleotide mix.
18\. The method of claim 1, wherein multiple different identifier sequences are detected in the cell or tissue sample, and each different identifier sequence is detected at a plurality of locations in the cell or tissue sample.
19\. The method of claim 1, wherein optical crowding is reduced compared to methods where the first and second identifier sequences are sequenced in the same series of sequencing cycles using the same sequencing primer.
20\. The method of claim 1, wherein the sequence in the arms of the first padlock probe is complementary to the first identifier sequence and the sequence in the arms of the second padlock probe is complementary to the second identifier sequence.

OFFICE ACTION:
Allowance notice
1.	The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
2.	This application is a CON of 18/589,354 02/27/2024, which is a CON of PCT/US2023/021925 filed on 05/11/2023, which claims benefit of 63/340,730 filed on 05/11/2022.
Information Disclosure Statement
3.	The information disclosure statement (IDS) submitted on 01/13/2025 has been considered by the examiner.  All the references filed there have been considered by the examiner.
It is in the examiner’s opinion that all the references cited in said IDS do not teach combination steps of instant claim 1.
Claim status
4.	In the claim listing filed on 10/24/2024 claims 1-20 are pending in this application.
Interview summary
5. 	Upon further search and other considerations the examiner contacted the representative Mr. Crawford to inform that claim 1 filed on 10/24/2024 contain extra space between 3 and '. During the interview the representative authorized to delete the extra space via examiner's amendment.
EXAMINER’S AMENDMENT
6.	Authorization for this examiner’s amendment was given in a telephonic interview with applicant’s representative Mr. Crawford on 01/24/2025.
Please arrange to delete extra space between 3 and ' in lines 5 and 7.
REASONS FOR ALLOWANCE
7.	The following is an examiner’s statement of reasons for allowance.
Cited arts: Pathak et al (WO 2005/061735 published July 07 2005) discloses a method of selectively amplifying a first strand or a second strand of a target nucleic acid molecule, comprising: contacting a padlock probe comprising a 5' end and a 3' end with a nucleic acid molecule, wherein the 5' end and the 3' end of the padlock probe specifically hybridize only to the first strand or to only the second strand of the target nucleic acid molecule; incubating the padlock probe and the nucleic acid molecule for a time sufficient to permit hybridization between the 5' end and the 3' end of the padlock probe and the target nucleic acid molecule; joining the 5' end and the 3' end of the padlock probe to one another, thereby generating a closed circularized padlock probe; amplifying the closed circularized padlock probe, thereby generating an amplicon; and amplifying the amplicon, thereby selectively generating amplified copies of the first strand or second strand of the target nucleic acid molecule and detecting
the amplified amplicon, wherein the method further comprises quantitating the
selectively generated amplified copies of the first strand or second strand of the
target nucleic acid molecule, wherein an amount of amplified copies is indicative of
an amount of first strand or second strand target nucleic acid molecule present in the sample (Pathak, claims 1 and 2).
Johnson et al (US 2015/0154352 published Jun. 4, 2025) disclose in Figs. 10 and 11 the components required for a single cell sequence linkage by padlock probes
combined with overlap extension polymerase chain reaction, according to one embodiment of the invention. FIG. 11 shows the complementary regions between a first padlock probe (a) and the first target nucleic acid ( c) and between a second padlock probe (b) and a second target nucleic acid ( d) in a single cell, according to one embodiment of the invention. The reaction components can be contained in a physical reaction container or an emulsion droplet (k). The first padlock probe (a) includes two separate regions that are complementary to the first target nucleic acid (c). The second padlock probe (b) includes two separate regions that are complementary
to a second target nucleic acid (d). A polymerase and a ligase are used (m) to amplify and ligate the gap between complementary regions of the padlock probes (a) and (b).
It is in the examiner’s opinion that although Pathak and Johnson teach some components and steps of claim 1, they do not teach combination of steps and components of instant claim 1.
Conclusion
8.	Claims 1-20 are allowed.
Any comments considered necessary by applicant must be submitted no later than the payment of the issue fee and, to avoid processing delays, should preferably accompany the issue fee.  Such submissions should be clearly labeled “Comments on Statement of Reasons for Allowance.”



CITED REFERENCE SUMMARIES: Ref US20150154352A1: Claims:
- 94 . (canceled)
A computer-implemented method for scoring a sample suspected of containing a heterogeneous mixture of target and background cells, comprising: a) obtaining a first variable corresponding to an average target sequence signal per cell in a substantially homogeneous population of target cells; b) obtaining a second variable corresponding to an average target sequence signal per cell in a substantially homogeneous population of background cells; c) obtaining a dataset obtained from a sample suspected of containing a heterogeneous mixture of target and background cells, wherein said dataset comprises quantitative sequence information derived from a set of individual nucleic acid molecules each comprising a target sequence linked to an identification sequence, wherein each of said identification sequences is associated with an individual droplet or reaction container comprising at least one cell from said sample; and d) inputting said first variable, said second variable, and said dataset into an interpretation function to determine a score that is indicative of the presence of at least one of said target cells within said individual droplet or reaction container.
a) obtaining a first variable corresponding to an average target sequence signal per cell in a substantially homogeneous population of target cells;
b) obtaining a second variable corresponding to an average target sequence signal per cell in a substantially homogeneous population of background cells;
c) obtaining a dataset obtained from a sample suspected of containing a heterogeneous mixture of target and background cells, wherein said dataset comprises quantitative sequence information derived from a set of individual nucleic acid molecules each comprising a target sequence linked to an identification sequence, wherein each of said identification sequences is associated with an individual droplet or reaction container comprising at least one cell from said sample; and
d) inputting said first variable, said second variable, and said dataset into an interpretation function to determine a score that is indicative of the presence of at least one of said target cells within said individual droplet or reaction container.
The method of claim 95 , wherein said identification sequence is an artificial barcode sequence.
The method of claim 95 , wherein said identification sequence is an endogenous variable sequence.
The method of claim 95 , wherein said target sequence is a gene.
The method of claim 95 , wherein said target sequence is present in fewer than 5% of the cells of said first dataset.
The method of claim 95 , wherein said target sequence is ligated to said identification sequence.
The method of claim 95 , wherein said score correlates to the presence or absence of a target sequence in one or more cells of said sample suspected of containing a heterogeneous mixture of target and background cells.
The method of claim 95 , wherein said quantitative sequence information is obtained from performing a sequencing reaction.
The method of claim 95 , wherein said quantitative sequence information is obtained from performing a quantitative polymerase chain reaction.
The method of claim 95 , wherein obtaining said first variable, said second variable, or said dataset comprises performing a sequencing reaction.
The method of claim 95 , wherein obtaining said first variable, said second variable, or said dataset comprises performing a quantitative polymerase chain reaction.
A kit for use in scoring a sample suspected of containing a heterogeneous mixture of target and background cells, comprising: a set of reagents comprising a plurality of reagents for obtaining a dataset from a sample suspected of containing a heterogeneous mixture of target and background cells, wherein said dataset comprises quantitative sequence information derived from a set of individual nucleic acid molecules each comprising a target sequence linked to an identification sequence, wherein each of said identification sequences is associated with an individual droplet or reaction container comprising at least one cell from said sample; and instructions for using said plurality of reagents to determine a score that is indicative of the presence of at least one of said target cells within said individual droplet or reaction container from said dataset, wherein said score is determined from an interpretation function, wherein said interpretation function comprises a first variable, a second variable, and said dataset, wherein said first variable corresponds to an average target sequence signal per cell in a substantially homogeneous population of target cells, and wherein said second variable corresponds to an average target sequence signal per cell in a substantially homogeneous population of background cells.
a set of reagents comprising a plurality of reagents for obtaining a dataset from a sample suspected of containing a heterogeneous mixture of target and background cells, wherein said dataset comprises quantitative sequence information derived from a set of individual nucleic acid molecules each comprising a target sequence linked to an identification sequence, wherein each of said identification sequences is associated with an individual droplet or reaction container comprising at least one cell from said sample; and
instructions for using said plurality of reagents to determine a score that is indicative of the presence of at least one of said target cells within said individual droplet or reaction container from said dataset, wherein said score is determined from an interpretation function, wherein said interpretation function comprises a first variable, a second variable, and said dataset, wherein said first variable corresponds to an average target sequence signal per cell in a substantially homogeneous population of target cells, and wherein said second variable corresponds to an average target sequence signal per cell in a substantially homogeneous population of background cells.
A computer-implemented method for scoring a first sample obtained from a first population of cells, comprising: obtaining a first dataset associated with a first sample obtained from a first population of cells, wherein said first dataset comprises quantitative sequence information derived from a first set of individual nucleic acid molecules each comprising a target sequence linked to an identification sequence from a set of N distinct identification sequences, and wherein each of said N distinct identification sequences is associated with an individual droplet or reaction container comprising a sample cell from which said first dataset was obtained; determining a first distribution comprising the number of said target sequences linked to each of said N distinct identification sequences; and analyzing said first distribution and a second distribution to determine a score predictive of the presence of a target cell within one or more cells of said first population of cells, wherein said second distribution is determined from a second dataset associated with a second sample obtained from a second population of cells, wherein said second dataset comprises quantitative sequence information derived from a second set of individual nucleic acid molecules each comprising said target sequence linked to an identification sequence from a set of Y distinct identification sequences, wherein each of said Y distinct identification sequences is associated with an individual droplet or reaction container comprising a sample cell from which said second dataset was obtained; and wherein said second distribution comprises the number of said target sequences linked to each of said Y distinct identification sequences.
obtaining a first dataset associated with a first sample obtained from a first population of cells, wherein said first dataset comprises quantitative sequence information derived from a first set of individual nucleic acid molecules each comprising a target sequence linked to an identification sequence from a set of N distinct identification sequences, and wherein each of said N distinct identification sequences is associated with an individual droplet or reaction container comprising a sample cell from which said first dataset was obtained;
determining a first distribution comprising the number of said target sequences linked to each of said N distinct identification sequences; and
analyzing said first distribution and a second distribution to determine a score predictive of the presence of a target cell within one or more cells of said first population of cells, wherein said second distribution is determined from a second dataset associated with a second sample obtained from a second population of cells, wherein said second dataset comprises quantitative sequence information derived from a second set of individual nucleic acid molecules each comprising said target sequence linked to an identification sequence from a set of Y distinct identification sequences, wherein each of said Y distinct identification sequences is associated with an individual droplet or reaction container comprising a sample cell from which said second dataset was obtained; and wherein said second distribution comprises the number of said target sequences linked to each of said Y distinct identification sequences.
The method of claim 107 , wherein said identification sequence is an artificial barcode sequence.
The method of claim 107 , wherein said identification sequence is an endogenous variable sequence.
The method of claim 107 , wherein said target sequence is an endogenous indicator sequence.
The method of claim 107 , wherein said target sequence is a gene.
The method of claim 107 , wherein said target sequence is present in fewer than 5% of the cells of said first dataset.
The method of claim 107 , wherein said target sequence is ligated to said identification sequence.
The method of claim 107 , wherein said score correlates to the presence or absence of a target cell in one or more cells of said first population of cells.
The method of claim 114 , wherein the presence of said target cell is indicative of an abnormality.
The method of claim 115 , wherein said abnormality is selected from the group consisting of a cancer, an inflammatory condition, a cardiovascular disease, an endocrine disease, an eye disease, a genetic disorder, an infectious disease, an intestinal disease, and a neurological disorder.
The method of claim 107 , wherein said target sequence comprises a sequence variation.
The method of claim 117 , wherein said sequence variation is a genetic mutation.
The method of claim 118 , wherein said genetic mutation is a mutation in a gene selected from the group consisting of: epidermal growth factor receptor (EGFR), phosphatase and tensin homolog (PTEN), tumor protein 53 (p53), MutS homolog 2 (MSH2), multiple endocrine neoplasia 1 (MEN1), adenomatous polyposis coli (APC), Fas receptor (FASR), retinoblastoma protein (Rb1), Janus kinase 2 (JAK2), (ETS)-like transcription factor 1 (ELK1), v-ets avian erythroblastosis virus E26 oncogene homolog 1 (ETS1), breast cancer 1 (BRCA1), breast cancer 2 (BRCA2), hepatocyte growth factor receptor (MET), ret protocooncogene (RET), V-erb-b2 erythroblastic leukemia viral oncogene homolog 2 (HER2), V-Kiras2 Kirsten rat sarcoma viral oncogene homolog (KRAS), B-cell lymphoma 2 (BCL2), Vmyc myelocytomatosis viral oncogene homolog (MYC), neurofibromatosis type 2 gene (NF2), v-myb myeloblastosis viral oncogene homolog (MYB), and mutS homolog 6 ( E. coli ) (MSH6).
The method of claim 107 , wherein N is at least 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , or 10 8 .
The method of claim 107 , wherein Y is at least 10 3 , 10 4 , 10 5 , 10 6 , 10 7 , or 10 8 .
The method of claim 107 , wherein N is the same as Y.
The method of claim 122 , wherein the sequences of said N distinct identification sequences are the same as the sequences of said Y distinct identification sequences.
The method of claim 107 , wherein said quantitative sequence information is obtained from performing a sequencing reaction.
The method of claim 107 , wherein said quantitative sequence information is obtained from performing a quantitative polymerase chain reaction.
Ref WO2005061735: Claims:
A method of selectively amplifying a first strand or a second strand of a target nucleic acid molecule, comprising: contacting a padlock probe comprising a 5' end and a 3' end with a nucleic acid molecule, wherein the 5' end and the 3' end of the padlock probe specifically hybridize only to the first strand or to only the second strand of the target nucleic acid molecule; incubating the padlock probe and the nucleic acid molecule for a time sufficient to permit hybridization between the 5' end and the 3' end of the padlock probe and the target nucleic acid molecule; joining the 5' end and the 3' end of the padlock probe to one another, thereby generating a closed circularized padlock probe; amplifying the closed circularized padlock probe, thereby generating an amplicon; and amplifying the amplicon, thereby selectively generating amplified copies of the first strand or second strand of the target nucleic acid molecule.
The method of claim 1, further comprising detecting the amplified amplicon.
The method of claim 1, wherein the method further comprises quantitating the selectively generated amplified copies of the first strand or second strand of the target nucleic acid molecule, wherein an amount of amplified copies is indicative of an amount of first strand or second strand target nucleic acid molecule present in the sample.
The method of claim 3, wherein the padlock probe is specific for the first strand and generates a tandem sequence complementary to the first sfrand, whereby a quantitation of the tandem sequence is proportional to a relative quantity of the first strand.
The method of claim 3, wherein the probe is specific for the second sfrand and generates a tandem sequence complementary to the second strand, whereby a  quantitation of the tandem sequence is proportional to a relative quantity of the second strand.
The method of claim 1, wherein amplifying the closed circularized padlock probe comprises using rolling circle amplification (RCA) or polymerase chain reaction
The method of claim 2, wherein detecting the amplified amplicon comprises using real-time quantitative PCR.
The method of claim 1, wherein the amplicon comprises a tandem sequence complementary to the closed circularized padlock probe.
The method of claim 1, wherein the method is a method of selectively amplifying a first strand and a second strand of a target nucleic acid molecule, wherein at least two padlock probes are incubated with the nucleic acid molecule for a time sufficient to permit hybridization between the 5' end and the 3' end of at least one of the padlock probes with the first strand of the target nucleic acid molecule and to permit hybridization between the 5' end and the 3' end of at least one of the padlock probes with the second strand of the target nucleic acid molecule, wherein the method permits simultaneous detection of the first strand and second strand of the target nucleic acid molecule.
The method of claim 1, wherein the method is a method of selectively amplifying a first strand or a second strand of at least two target nucleic acid molecules, wherein at least two padlock probes are incubated with the nucleic acid molecule for a time sufficient to permit hybridization between the 5' end and the 3' end of at least one of the padlock probes with at least one target nucleic acid molecule and to permit hybridization between the 5' end and the 3' end of at least one of the padlock probes with another target nucleic acid molecule, wherein the method permits simultaneous detection of at least two different first strands and at least two different second strands of the target nucleic acid molecules.
The method of claim 2, wherein detecting the amplicon comprises incubating the amplicon with a primer complementary to the 5' end and the 3' end of the padlock probe, wherein the primer comprises a reporter fluorophore and a quencher.
The method of claim 11 , wherein the reporter fluorophore comprises 6-carboxy- fluorescein (FAM), and the quencher comprises 6-carboxy-tetramethylrhodamine (TAMRA).
The method of claim 1, wherein joining the 5' end and the 3' end of the padlock probe, comprises ligating the 5' end and the 3' end of the padlock probe together.
The method of claim 1, wherein when the 5' end and the 3' end of the padlock probe specifically hybridize only to the first strand or to only the second sfrand of the target nucleic acid molecule, a gap is present between the 5' end and the 3' end of the padlock probe.
The method of claim 14, wherein the gap is no more than about 20 nucleotides.
The method of claim 14, wherein joining the 5' end and the 3' end of the padlock probe to one another comprises filling in the gap and ligating the 5' end and the 3' end of the padlock probe together.
The method of claim 1, wherein the padlock probe comprises a spacer region that includes a detectable sequence.
The method of claim 17, wherein the detectable sequence is complementary to a primer sequence.
The method of claim 18, wherein the primer sequence comprises a fluorophore.
The method of claim 2, further comprising: providing a second padlock probe specific for the second strand;  contacting the second padlock probe comprising a 5' end and a 3' end with the nucleic acid molecule; incubating the second padlock probe and the nucleic acid molecule for a time sufficient to permit hybridization between the 5' end and the 3' end of the second padlock probe and the target nucleic acid molecule; ligating the 5' end and the 3' end of the second padlock probe, thereby generating a second closed circularized padlock probe; amplifying the second closed circularized padlock probe, thereby generating a second amplicon; amplifying the second amplicon; and quantitating the second amplicon, whereby a quantity of the first and the second amplicon provides a relative indication of a relative quantity of the first strand and second strand.
The method of claim 20, wherein the first and second padlock probe are contacted and incubated with the with the nucleic acid molecule simultaneously, wherein ligating the 5' end and the 3' end of the first padlock probe and second padlock probe occurs simultaneously, wherein the first closed circularized padlock probe and the second closed circularized padlock probe are amplified simultaneously, wherein the first amplicon and the second amplicon are amplified simultaneously, and wherein the first amplicon and the second amplicon are quantified simultaneously.
The method of claim 1, wherein the target nucleic acid molecule comprises a viral nucleic acid molecule.
The method of claim 22, wherein the first sfrand is a plus-sfrand and the second strand is a minus strand.
The method of claim 22, wherein the viral nucleic acid molecule comprises a Human Immunodeficiency Virus (HIV) nucleic acid molecule.
The method of claim 24, wherein the method is a method of selectively amplifying a plus-strand of HIV, and the padlock probe comprises any of SEQ ID NOS: 7-12.
The method of claim 24, wherein the method is a method of selectively amplifying a minus-strand of HIN, and the padlock probe comprises any of SEQ ID ÎOS: 1-6.
The method of 1, wherein the padlock probe comprises any of SEQ ID ÎOS: 1- 12.
The method of claim 1, wherein the first strand is a leading-strand and the second strand is a lagging-strand.
A method of selectively amplifying and quantitating a first sfrand or a second strand of a double-stranded nucleic acid molecule, comprising: contacting a padlock probe comprising a 5' end and a 3' end with a nucleic acid molecule, wherein the padlock probe is specific for the first strand or the second stra
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