DETAILED ACTION
Notice of Pre-AIA or AIA Status
1. The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Claim Objections
2. Claim 7 is objected to because of the following informalities: “method of any of claim 1” as recited in line 1 should be changed to “method of . Appropriate correction is required.
Claim Rejections - 35 USC § 112
3. The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
4. Claims 1-24 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor, or for pre-AIA the applicant regards as the invention.
Claim 1 recites a step of “stimulating light emissions from the first and second labeled primers, wherein an amplitude of the signal generated by the first labeled primers is greater than an amplitude of the signal generated by the second labeled primers” followed by a step of “identifying the labeled nucleobases added to the first primers and the second primers based on the amplitude of the signal generated by the labeled nucleobases” (emphasis provided). First, it is not clear whether “the labeled nucleobases” as recited in the “identifying” step refers to all the “labeled nucleobases” used in the “extending” step (where only some of the “labeled nucleobases” would be incorporated into the “first primers” and the “second primers”) or only the incorporated “labeled nucleobases”. Second, since there would be a signal generated by the “labeled nucleobases” incorporated into the “first primers” and another signal generated by the “labeled nucleobases” incorporated into the “second primers”, does “the signal generated by the labeled nucleobases” (as recited in the “identifying” step) refers to one of those two signals, both of those two signals, or a combined signal from those two signals? Claims 2-24, each of which depends from claim 1, are also rejected for the same reasons.
In the interest of compact prosecution and for purposes of current examination, the “identifying” step in claim 1 is being interpreted as “identifying the labeled nucleobases added to the first primers based on the amplitude of the signal generated by the labeled nucleobases incorporated into the first primers and identifying the labeled nucleobases added to the second primers based on the amplitude of the signal generated by the labeled nucleobases incorporated into the second primers”.
Claim Rejections - 35 USC § 102
5. In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
6. The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
7. Claims 1, 3-7, 13, 16-19, 22-23, 25-32, 35-38 and 41 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Smith (WO 2007/010252 A1).
Regarding claim 1
Smith teaches, throughout the whole document, a method of identifying nucleobases in a template polynucleotide, comprising: providing a substrate (e.g., solid support) comprising a plurality of double stranded template polynucleotides in a cluster, wherein each double stranded template polynucleotide comprises a first strand and a second strand (see page 6, lines 8-13; page 7, line 15 – page 8, line 7; page 30, line 27 – page 31, line 11; Figure 1); contacting the plurality of double stranded template polynucleotides with first primers which bind to the first strand and second primers which bind to the second strand (see page 6, lines 18-20 and 30-32; Figure 1); extending the first primers and the second primers by contacting the cluster with labeled nucleobases (e.g., fluorescently labeled nucleotides. See page 36, lines 22-25; page 57, lines 6-9) to form first labeled primers and second labeled primers (see page 6, lines 21-24; page 7, lines 1-4; page 33, line 27 – page 35, line 18; Figure 1); stimulating (e.g., stimulating by using laser light) light emissions from the first and second labeled primers (see paragraph bridging pages 36-37), wherein an amplitude of the signal generated by the first labeled primers is greater than an amplitude of the signal generated by the second labeled primers (i.e., since the two strands are complementary rather than identical, the nucleotides incorporated into the first labeled primers and the second labeled primers are different. Because the different fluorescent labels on different nucleotides (see page 36, lines 22-25; page 57, lines 6-9) have different peak emission wavelengths and different extinction coefficients, the amplitudes of the signals generated by the first and second labeled primers would also be different, meaning one is greater than the other. Note that the “first” or “second” designation for the two strands (and thus the two labeled primers) is arbitrary, meaning that the “first” strand/primer used in Smith’s method may be designated as either “first” or “second” strand/primer to meet the claim.); and identifying the labeled nucleobases added to the first primers based on the amplitude of the signal generated by the labeled nucleobases incorporated into the first primers and identifying the labeled nucleobases added to the second primers based on the amplitude of the signal generated by the labeled nucleobases incorporated into the second primers (see page 36, line 22 – page 37, line 3; page 57, lines 6-27).
Regarding claim 3
The method according to Smith, wherein the signal generated by the first labeled primers and the signal generated by the second labeled primers are emitted from the same region or substantially overlapping regions of the substrate (see page 7, line 15 – page 8, line 7; page 30, line 27 – page 31, line 11; page 36, line 22 – page 37, line 3; page 57, lines 6-27; Figure 1).
Regarding claim 4
The method according to Smith, wherein either the first strand or the second strand of each double stranded template polynucleotide is attached to the substrate (see Figure 1).
Regarding claim 5
The method according to Smith, wherein the plurality of double stranded template polynucleotides in the cluster are generated by a bridge amplification process (see paragraph bridging pages 46-47; Figure 1).
Regarding claim 6
The method according to Smith, wherein the substrate comprises a plurality of clusters of nucleic acids, the clusters being randomly distributed on the substrate (see paragraphs bridging pages 7-8; page 30, line 27 – page 31, line 11; Figure 1).
Regarding claim 7
The method according to Smith, wherein the amplitude of the signal generated by the first labeled primers would necessarily correspond with a (first) quantity of the first labeled primers in the cluster, and wherein the amplitude of the signal generated by the second labeled primers would necessarily correspond with a (second) quantity of the second labeled primers in the cluster.
Regarding claim 13
The method according to Smith, wherein extending the first primers and extending the second primers are catalyzed by a strand-displacing polymerase (e.g., 9°N polymerase) (see page 57, lines 6-13).
Regarding claim 16
The method according to Smith, comprising: detecting the signal generated by the first labeled primers in a first range of optical frequencies and a second range of optical frequencies (e.g., in different ranges of optical frequencies corresponding to the different fluorescent labels of the different fluorescently labeled nucleotides used in the method); and detecting the signal generated by the second labeled primers in the first range of optical frequencies and the second range of optical frequencies (e.g., in different ranges of optical frequencies corresponding to the different fluorescent labels of the different fluorescently labeled nucleotides used in the method), wherein the first range of optical frequencies and the second range of optical frequencies are not identical (see page 36, line 22 – page 37, line 3; page 57, lines 6-27).
Regarding claim 17
The method according to Smith, comprising: acquiring a first fluorescent image of the cluster in a first range of optical frequencies; acquiring a second fluorescent image of the cluster in a second range of optical frequencies, wherein the first range of optical frequencies and the second range of optical frequencies are not identical (e.g., different ranges of optical frequencies due to the different fluorescent labels of the different fluorescently labeled nucleotides used in the method); and obtaining the signals generated by the first and second labeled primers by extracting fluorescence intensities from the first and second fluorescent images of the cluster (see page 36, line 22 – page 37, line 3; page 57, lines 6-27).
Regarding claim 18
The method according to Smith, comprising extracting fluorescence intensities from the first and second fluorescent images of the same region or substantially overlapping regions of the substrate (see page 36, line 22 – page 37, line 3; page 57, lines 6-27; Figure 1).
Regarding claim 19
The method according to Smith, wherein identifying the labeled nucleobases added to the first primers and the second primers is based on a combination of the extracted fluorescence intensities from the first and second fluorescent images (see page 36, line 22 – page 37, line 3; page 57, lines 6-27).
Regarding claim 22
The method according to Smith, comprising stimulating fluorescent emissions from the first labeled primers and second labeled primers in the cluster with light at a predetermined optical frequency (e.g., light at a predetermined optical frequency corresponding to the excitation wavelength of the fluorescent label of each of the fluorescently labeled nucleotides used in the method) (see page 36, line 22 – page 37, line 3; page 57, lines 6-27).
Regarding claim 23
The method according to Smith, comprising stimulating fluorescent emissions from the first labeled primers and second labeled primers in the cluster with light at two predetermined optical frequencies (e.g., light at two predetermined optical frequencies corresponding to the excitation wavelengths of the fluorescent labels of any two of the fluorescently labeled nucleotides used in the method) (see page 36, line 22 – page 37, line 3; page 57, lines 6-27).
Regarding claim 25
Smith teaches, throughout the whole document, a method of determining the sequence of a template polynucleotide, the method comprising: hybridizing a first primer to the template polynucleotide and a second primer to the reverse complement of the template polynucleotide, wherein the template polynucleotide and the reverse complement of the template polynucleotide are at substantially overlapping regions of a substrate (see page 6, lines 3-32; Figure 1); extending the first primer with a first labeled nucleotide analog (e.g., fluorescently labeled nucleotide analog. See page 36, lines 22-25; page 57, lines 6-9) and extending the second primer with a second labeled nucleotide analog (see page 6, lines 21-24; page 7, lines 1-4; page 33, line 27 – page 35, line 18; Figure 1); stimulating (e.g., stimulating by using laser light) light emissions from the first and second labeled nucleotide analogs (see paragraph bridging pages 36-37); and determining the sequence of nucleotides in the template polynucleotide and the reverse complement of the template polynucleotide by capturing the light emissions nucleobases (see page 36, line 22 – page 37, line 3; page 57, lines 6-27).
Regarding claim 26
The method according to Smith, wherein the template polynucleotide and the reverse complement of the template polynucleotide are part of a cluster of identical copies of the template polynucleotide and identical copies the reverse complement of the template polynucleotide (see page 7, lines 15-27; page 30, line 27 – page 31, line 11; Figure 1).
Regarding claim 27
The method according to Smith, wherein the cluster of identical copies of the template polynucleotide and identical copies the reverse complement of the template polynucleotide is generated by bridge amplification (see paragraph bridging pages 46-47; Figure 1).
Regarding claims 28-29
The method according to Smith, wherein the identical copies of the template polynucleotide have an end attached to the substrate by a first grafting oligonucleotide, and wherein the identical copies of the reverse complement of the template polynucleotide have an end attached to the substrate by a second grafting oligonucleotide (see page 38, lines 1-32).
Regarding claim 30
The method according to Smith, wherein at least a portion of the reverse complement of the template polynucleotide is hybridized with a portion of the template polynucleotide (see Figure 1).
Regarding claim 31
The method according to Smith, wherein the first primer is part of a first population of first primers hybridized to identical copies of the template polynucleotide, and wherein the second primer is part of a second population of second primers hybridized to identical copies of the reverse complement of the template polynucleotide (see page 6, lines 18-32; claim 1).
Regarding claim 32
The method according to Smith, wherein determining the sequence of nucleotides comprises: receiving a first signal emitted at a first amplitude from the first population of first primers; receiving a second signal emitted at a second amplitude from the second population of second primers; and identifying a nucleobase hybridized to the template polynucleotide and a nucleobase hybridized to the reverse complement of the template polynucleotide based on a combination of the first and second signals (see page 36, line 22 – page 37, line 3; page 57, lines 6-27).
Regarding claim 35
The method according to Smith, wherein the first population of first primers have an unblocked 3’-end (so that they can be extended) (see page 6, lines 18-24; Figure 1).
Regarding claim 36
The method according to Smith, wherein the first primer and the second primer are hybridized to the template polynucleotide and the reverse complement of the template polynucleotide, respectively, in the same reaction step (when the sequencing of both strands is considered as a large step) (see page 6, lines 3-32; Figure 1).
Regarding claim 37
The method according to Smith, wherein extending the first primer with the first labeled nucleotide analog and extending the second primer with the second labeled nucleotide analog are performed in the same reaction step (when the sequencing of both strands is considered as a large step) (see page 6, lines 21-24; page 7, lines 1-4; page 33, line 27 – page 35, line 18; Figure 1).
Regarding claim 38
The method according to Smith, wherein the first labeled nucleotide analog and the second labeled nucleotide analog are hybridized to the template polynucleotide and the reverse complement of the template polynucleotide, respectively, in the same reaction step (when the sequencing of both strands is considered as a large step) (see page 6, line 18 – page 7, line 4; page 33, line 27 – page 35, line 18).
Regarding claim 41
The method according to Smith, wherein extending the first primer and extending the second primer are catalyzed by a strand-displacing polymerase (e.g., 9°N polymerase) (see page 57, lines 6-13).
8. Claims 1, 3-7, 10-19, 22-23, 25-32 and 35-43 are rejected under 35 U.S.C. 102(a)(2) as being anticipated by Witters et al. (WO 2022/170212 A1).
Regarding claim 1
Witters et al. teach, throughout the whole document, a method of identifying nucleobases in a template polynucleotide, comprising: providing a substrate comprising a plurality of double stranded template polynucleotides in a cluster, wherein each double stranded template polynucleotide comprises a first strand and a second strand (see paragraph [0012]; Figures 1A and 1B); contacting the plurality of double stranded template polynucleotides with first primers which bind to the first strand and second primers which bind to the second strand (see paragraphs [0006]-[0012]; claims 1 and 26; Figures 1A and 1B); extending the first primers and the second primers by contacting the cluster with labeled nucleobases (e.g., fluorescently labeled nucleotides. See paragraphs [0208]-[0209] and [0218]) to form first labeled primers and second labeled primers (see paragraphs [0006]-[0012]; claims 1 and 26; Figures 1A and 1B); stimulating (e.g., stimulating by using laser light) light emissions from the first and second labeled primers (see paragraph [0218]), wherein an amplitude of the signal generated by the first labeled primers is greater than an amplitude of the signal generated by the second labeled primers (i.e., since the two strands are complementary rather than identical, the nucleotides incorporated into the first labeled primers and the second labeled primers are different. Because the different fluorescent labels on different nucleotides (see paragraph [0218]) have different peak emission wavelengths and different extinction coefficients, the amplitudes of the signals generated by the first and second labeled primers would also be different, meaning one is greater than the other. Note that the “first” or “second” designation for the two strands (and thus the two labeled primers) is arbitrary, meaning that the “first” strand/primer used in the method of Witters et al. may be designated as either “first” or “second” strand/primer to meet the claim.); and identifying the labeled nucleobases added to the first primers based on the amplitude of the signal generated by the labeled nucleobases incorporated into the first primers and identifying the labeled nucleobases added to the second primers based on the amplitude of the signal generated by the labeled nucleobases incorporated into the second primers (see paragraphs [0006]-[0012], [0074], [0209] and [0218]; Figures 1A and 1B).
Regarding claim 3
The method according to Witters et al., wherein the signal generated by the first labeled primers and the signal generated by the second labeled primers are emitted from the same region or substantially overlapping regions of the substrate (see paragraphs [0006]-[0012; Figures 1A and 1B).
Regarding claim 4
The method according to Witters et al., wherein either the first strand or the second strand of each double stranded template polynucleotide is attached to the substrate (see paragraph [0012]; Figures 1A and 1B).
Regarding claim 5
The method according to Witters et al., wherein the plurality of double stranded template polynucleotides in the cluster are generated by a bridge amplification process (see paragraphs [0195], [0292] and [0350]).
Regarding claim 6
The method according to Witters et al., wherein the substrate comprises a plurality of clusters of nucleic acids, the clusters being randomly distributed on the substrate (see paragraph [0012]; Figures 1A and 1B).
Regarding claim 7
The method according to Witters et al., wherein the amplitude of the signal generated by the first labeled primers would necessarily correspond with a (first) quantity of the first labeled primers in the cluster, and wherein the amplitude of the signal generated by the second labeled primers would necessarily correspond with a (second) quantity of the second labeled primers in the cluster.
Regarding claims 10-11
The method according to Witters et al., wherein the first primers and the second primers are formed of a locked nucleic acid (LNA) or a peptide nucleic acid (PNA) (see paragraphs [0037], [0105], [0125]-[0127] and [0155]-[0157]).
Regarding claim 12
The method according to Witters et al., comprising contacting the plurality of double stranded template polynucleotides with a RecA-like protein or a non-nicking CRISPR-associated protein to facilitate binding of the plurality of double stranded template polynucleotides with the first primers and the second primers (see paragraphs [0184], [0256] and [0366]).
Regarding claims 13-14
The method according to Witters et al., wherein extending the first primer and extending the second primer are catalyzed by a strand-displacing polymerase, wherein the strand-displacing polymerase comprises Klenow fragment, phi29 DNA polymerase, Bsm DNA polymerase, Bst DNA polymerase, or conserved mutations thereof (see paragraph [0122]).
Regarding claim 15
The method according to Witters et al., comprising contacting the plurality of double stranded template polynucleotides with a helicase, a single-stranded DNA binding protein, or a mixture of oligonucleotides having random sequences, to partially separate the first strand and the second strand of each double stranded template polynucleotide (see paragraphs [0059], [0061] and [0179]).
Regarding claim 16
The method according to Witters et al., comprising: detecting the signal generated by the first labeled primers in a first range of optical frequencies and a second range of optical frequencies (e.g., in different ranges of optical frequencies corresponding to the different fluorescent labels of the different fluorescently labeled nucleotides used in the method); and detecting the signal generated by the second labeled primers in the first range of optical frequencies and the second range of optical frequencies (e.g., in different ranges of optical frequencies corresponding to the different fluorescent labels of the different fluorescently labeled nucleotides used in the method), wherein the first range of optical frequencies and the second range of optical frequencies are not identical (see paragraphs [0006]-[0012], [0074], [0209] and [0218]).
Regarding claim 17
The method according to Witters et al., comprising: acquiring a first fluorescent image of the cluster in a first range of optical frequencies; acquiring a second fluorescent image of the cluster in a second range of optical frequencies, wherein the first range of optical frequencies and the second range of optical frequencies are not identical (e.g., different ranges of optical frequencies due to the different fluorescent labels of the different fluorescently labeled nucleotides used in the method); and obtaining the signals generated by the first and second labeled primers by extracting fluorescence intensities from the first and second fluorescent images of the cluster (see paragraphs [0006]-[0012], [0074], [0209] and [0218]).
Regarding claim 18
The method according to Witters et al., comprising extracting fluorescence intensities from the first and second fluorescent images of the same region or substantially overlapping regions of the substrate (see paragraphs [0006]-[0012], [0074], [0209] and [0218]; Figures 1A and 1B).
Regarding claim 19
The method according to Witters et al., wherein identifying the labeled nucleobases added to the first primers and the second primers is based on a combination of the extracted fluorescence intensities from the first and second fluorescent images (see paragraphs [0006]-[0012], [0074], [0209] and [0218]).
Regarding claim 22
The method according to Witters et al., comprising stimulating fluorescent emissions from the first labeled primers and second labeled primers in the cluster with light at a predetermined optical frequency (e.g., light at a predetermined optical frequency corresponding to the excitation wavelength of the fluorescent label of each of the fluorescently labeled nucleotides used in the method) (see paragraphs [0006]-[0012], [0074], [0209] and [0218]).
Regarding claim 23
The method according to Witters et al., comprising stimulating fluorescent emissions from the first labeled primers and second labeled primers in the cluster with light at two predetermined optical frequencies (e.g., light at two predetermined optical frequencies corresponding to the excitation wavelengths of the fluorescent labels of any two of the fluorescently labeled nucleotides used in the method) (see paragraphs [0006]-[0012], [0074], [0209] and [0218]).
Regarding claim 25
Witters et al. teach, throughout the whole document, a method of determining the sequence of a template polynucleotide, the method comprising: hybridizing a first primer (e.g., first sequencing primer) to the template polynucleotide and a second primer (e.g., second sequencing primer) to the reverse complement of the template polynucleotide, wherein the template polynucleotide and the reverse complement of the template polynucleotide are at substantially overlapping regions of a substrate (see paragraphs [0006]-[0012]; claims 1 and 26; Figures 1A and 1B); extending the first primer with a first labeled nucleotide analog (e.g., fluorescently labeled nucleotide analog. See paragraphs [0208]-[0209] and [0218]) and extending the second primer with a second labeled nucleotide analog (see paragraphs [0006]-[0012]; claims 1 and 26; Figures 1A and 1B); stimulating (e.g., stimulating by using laser light) light emissions from the first and second labeled nucleotide analogs (see paragraph [0218]); and determining the sequence of nucleotides in the template polynucleotide and the reverse complement of the template polynucleotide by capturing the light emissions (see paragraphs [0006]-[0012], [0074], [0209] and [0218]; Figures 1A and 1B).
Regarding claim 26
The method according to Witters et al., wherein the template polynucleotide and the reverse complement of the template polynucleotide are part of a cluster of identical copies of the template polynucleotide and identical copies the reverse complement of the template polynucleotide (see paragraph [0012]).
Regarding claim 27
The method according to Witters et al., wherein the cluster of identical copies of the template polynucleotide and identical copies the reverse complement of the template polynucleotide is generated by bridge amplification (see paragraphs [0195], [0292] and [0350]).
Regarding claims 28-29
The method according to Witters et al., wherein the identical copies of the template polynucleotide have an end attached to the substrate by a first grafting oligonucleotide, and wherein the identical copies of the reverse complement of the template polynucleotide have an end attached to the substrate by a second grafting oligonucleotide (see paragraphs [0007] and [0111]-[0112]).
Regarding claim 30
The method according to Witters et al., wherein at least a portion of the reverse complement of the template polynucleotide is hybridized with a portion of the template polynucleotide (see Figures 1A and 1B).
Regarding claim 31
The method according to Witters et al., wherein the first primer is part of a first population of first primers hybridized to identical copies of the template polynucleotide, and wherein the second primer is part of a second population of second primers hybridized to identical copies of the reverse complement of the template polynucleotide (see paragraph [0012]; Figures 1A and 1B).
Regarding claim 32
The method according to Witters et al., wherein determining the sequence of nucleotides comprises: receiving a first signal emitted at a first amplitude from the first population of first primers; receiving a second signal emitted at a second amplitude from the second population of second primers; and identifying a nucleobase hybridized to the template polynucleotide and a nucleobase hybridized to the reverse complement of the template polynucleotide based on a combination of the first and second signals (see paragraphs [0006]-[0012], [0074], [0209] and [0218]; Figures 1A and 1B).
Regarding claim 35
The method according to Witters et al., wherein the first population of first primers have an unblocked 3’-end (so that they can be extended) (see Figures 1A and 1B).
Regarding claim 36
The method according to Witters et al., wherein the first primer and the second primer are hybridized to the template polynucleotide and the reverse complement of the template polynucleotide, respectively, in the same reaction step (when the sequencing of both strands is considered as a large step) (see paragraphs [0006]-[0012]; claims 1 and 26; Figures 1A and 1B).
Regarding claim 37
The method according to Witters et al., wherein extending the first primer with the first labeled nucleotide analog and extending the second primer with the second labeled nucleotide analog are performed in the same reaction step (when the sequencing of both strands is considered as a large step) (see paragraphs [0006]-[0012]; claims 1 and 26; Figures 1A and 1B).
Regarding claim 38
The method according to Witters et al., wherein the first labeled nucleotide analog and the second labeled nucleotide analog are hybridized to the template polynucleotide and the reverse complement of the template polynucleotide, respectively, in the same reaction step (when the sequencing of both strands is considered as a large step) (see paragraphs [0006]-[0012], [0208]-[0209] and [0218]; claims 1 and 26; Figures 1A and 1B).
Regarding claim 39
The method according to Witters et al., wherein the first primer and/or the second primer comprises a locked nucleic acid (LNA) or a peptide nucleic acid (PNA) (see paragraphs [0037], [0105], [0125]-[0127] and [0155]-[0157]).
Regarding claim 40
The method according to Witters et al., wherein hybridizing the first primer to the template polynucleotide and the second primer to the reverse complement of the template polynucleotide is facilitated by the presence of a RecA-like protein or a non-nicking CRISPR-associated protein (see paragraphs [0184], [0256] and [0366]).
Regarding claims 41-42
The method according to Witters et al., wherein extending the first primer and extending the second primer are catalyzed by a strand-displacing polymerase, wherein the strand-displacing polymerase comprises Klenow fragment, phi29 DNA polymerase, Bsm DNA polymerase, Bst DNA polymerase, or conserved mutations thereof (see paragraph [0122]).
Regarding claim 43
The method according to Witters et al., wherein the template polynucleotide and the reverse complement of the template polynucleotide are at least partially separated by the presence of a helicase, a single-stranded DNA binding protein, or a mixture of oligonucleotides having random sequences (see paragraphs [0059], [0061] and [0179]).
Conclusion
9. No claim is allowed.
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/KAIJIANG ZHANG/Primary Examiner, Art Unit 1684