Office Action 15: Notice of Allowance (2025-03-26)

CLAIMS:
Amendments to the claims:

Please amend the claims as follows.
1\. (Currently Amended) A method for droplet formation for sample preparation for sequencing, the method comprising:providing, via at least one inlet channel on a microfluidic substrate, an aqueous sample fluid comprising nucleic acid and at least one primer;providing an immiscible first carrier fluid t-t-enSVG {EMBED HERE} 0.1270.173uppe;appluelSVG {EMBED HERE} 0.0870.143gried in \[\[to\]\] a main channel on the microfluidic substrate;
 producing a plurality of droplets comprising the aqueous sample fluid at a junction between \[\[in\]\] the main channel and the at least one inlet channel;
copying at least a portion of the nucleic acid within one droplet of the plurality of droplets by extending the at least one primer to generate an extension ampeatien product; and
breaking the plurality of droplets by introducing a droplet-destabilizing second fluid comprising a destabilizing surfactant, thereby recovering the aqueous sample fluid including the extension product from the plurality of droplets.

2-3. (Cancelled)

4\. (Currently Amended) The method of claim 1, wherein the aqueous sample fluid comprises one or more cells hmpleelatieentinuequeehase.

5-11. (Cancelled)610

Application No. 15/480,739 12. (Currently Amended) The method of claim 1, further comprising subjecting the extension product to an amplification reaction.

13\. (Currently Amended) The method of claim 12, wherein the amplification reaction is a PCR reaction .

14-20. (Cancelled)1519

21\. (Currently Amended) The method of claim 1, wherein \[\[the\]\] at least one oil supply line provides a constant flow of the first carrier fluid into a nozzle while the plurality of droplets are formed.

22-30. (Cancelled)2329

31\. (Previously Presented) The method of claim 1, wherein the first carrier fluid comprises an oil.

32\. (Currently Amended) The method of claim 31, wherein the oil comprises a fluorocarbon oil or a silicone oil.

33\. (Previously Presented) The method of claim 31, wherein the oil comprises a surfactant.

34\. (Previously Presented) The method of claim 33, wherein the surfactant is a fluorosurfactant.

35-38. (Cancelled)3637

39\. (Currently Amended) The method of claim \[\[13\]\]1, wherein the microfluidic substrate is a microfluidic chip thadea\-aneees.
40\. (Currently Amended) The method of claim \[\[39\]\]1, wherein the aqueous sample fluid comprises one or more beads .

41\. (Cancelled)

42\. (Currently Amended) The method of claim \[\[4\]\] 1, further comprising centrifuging the plurality of droplets and the droplet-destabilizing second fluid to promote complete coalescence of the plurality of droplets.

43\. (Currently Amended) The method of claim \[\[4\]\] 1, wherein the destabilizing surfactant comprises a perfluorinated alcohol.

44\. (New) The method of claim 43, wherein the perfluorinated alcohol comprises 1H,1H,2H,2H-Perfluoro-1-octanol.

45\. (New) The method of claim 1, wherein the copying step uses a reverse transcriptase to produce complementary DNA.

46\. (New) A sample preparation method comprising:flowing, via an inlet channel on a microfluidic substrate, an aqueous sample fluid comprising nucleic acid to a junction between the inlet channel and a main channel having an immiscible carrier fluid therein;
producing a plurality of droplets comprising the aqueous sample fluid surrounded by the immiscible carrier fluid in the main channel;
copying at least a portion of the nucleic acid within one droplet of the plurality of droplets by extending a primer to generate an extension product; and
recovering the aqueous sample fluid including the extension product from the plurality of droplets by introducing a droplet-destabilizing second fluid comprising a perfluorinated alcohol.

Application No. 15/480,739 47. (New) The method of claim 46, wherein the perfluorinated alcohol comprises 1H,1 H,2H,2H-Perfluoro- 1 -octanol.

48\. (New) The method of claim 46, wherein the copying step uses a reverse transcriptase to produce complementary DNA.

49\. (New) The method of claim 46, wherein the aqueous fluid sample comprises one or more cells.

50\. (New) The method of claim 46, wherein the aqueous fluid sample comprises one or more beads.

OFFICE ACTION:
DETAILED ACTION
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection.  Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.  Applicant's submission filed on January 16, 2025 has been entered.
Preliminary Remark
Claims 2, 3, 5-11, 14-20, 22-30, 35-38, and 41 are canceled.
Claims 44-50 are new.
Claim Rejections - 35 USC § 103
The rejection of claims 1, 4, 12, 13, 21, 31, 39, and 40 under pre-AIA  35 U.S.C. 103(a) as being unpatentable over Cho et al. (EP 1 574 5861 A2, published 2005) in view of Kumacheva et al. (WO 2005/103106 A1, published November 2005, priority April 2004), and Dressman et al. (WO 2005/010145 A2, published February 2005; IDS ref), priority July 2003), made in the Office Action mailed on October 21, 2024 is withdrawn in view of the Amendment received on January 16, 2025.
The rejection of claims 32-34 under pre-AIA  35 U.S.C. 103(a) as being unpatentable over Cho et al. (EP 1 574 5862 A2, published 2005) in view of Kumacheva et al. (WO 2005/103106 A1, published November 2005, priority April 2004), and Dressman et al. (WO 2005/010145 A2, published February 2005; IDS ref), priority July 2003), as applied to claims 1, 4, 12, 13, 21, 31, 39, and 40 above, and further in view of Dorfman et al. (Analytical Chemistry, 2005, vol. 77, pages 3700-3704), made in the Office Action mailed on October 21, 2024 is withdrawn in view of the Amendment received on January 16, 2025.
Conclusion
Claims are free of prior art.
The prior art does not teach or suggest a method involving the mixing of droplets of the from a first carrier fluid (i.e., immiscible to aqueous sample fluid, see claim 1) with a droplet-destabilizing second fluid, wherein said destabilizing second fluid comprises an oil comprising a destabilizing surfactant, said surfactant comprising perfluorinated alcohol.
Instant application discloses that the use of oil comprising a destabilizing surfactant result in a gentle manner so as to minimize potential damage to the contents of the droplets:
“present invention proposes methods for recovering aqueous phase components from aqueous emulsions that have been collected on a microfluidic device in a minimum number of steps and in a gentle manner” (section [0279])
“a stable aqueous sample droplet emulsion containing phase components in a continuous phase carrier fluid is allowed to cream to the top of the continuous phase carrier oil … aqueous emulsion rises to the top or separates from the continuous phase fluid being greater than that of the aqueous phase emulsion … The creamed emulsion is then placed onto a second continuous phase carrier fluid which contains a de-stabilizing surfactant … the aqueous emulsion begins to coalesce … can now be removed” (sections [0280] and [0281])
Inquiries

APPLICANT REBUTTAL: Remarks
With entry of this amendment, claims 1, 4, 12, 13, 21, 31-34, 39, 40, and 42-50 are
pending, with claims 1 and 46 being the only independent claims. Claims 1, 4, 12, 13, 21, 32, 39, 40, 42, and 43 are amended. Claims 44-50 are new. Support for the amendments may be found throughout the as-filed application including, for example, at least at page 39, lines 20-23; page 100, lines 9-11; page 20, line 1; page 25, lines 11-12; page 22, lines 8-10; page 7, line 7; page 63, lines 16-19; page 25, lines 7-9; page 70, lines 4-6; page 33, lines 29-30; and page 91, lines 25-26.
No new matter is added.
Claims Free of the Prior Art
The Office Action, at page 14, states that claims 41-43 are free of prior art, but are
objected to for being dependent on a rejected base claim. The Office Action further states that, "The prior art does not teach or suggest a method involving the mixing of droplets of the from (sic) a first carrier fluid (i.e., immiscible to aqueous sample fluid, see claim 1) with a droplet- destabilizing second fluid, wherein said destabilizing second fluid comprises an oil comprising a destabilizing surfactant (claim 41), said surfactant comprising perfluorinated alcohol."
Applicant has amended the claims so that each of the independent claims recites subject matter from at least one of claims 41-43, which were found to be free of the prior art. Applicant submits that, with entry of this amendment, the independent claims-and thus all of the claims- are free of the prior art. Applicant thanks the Examiner for the review and analysis.
Rejections under 35 U.S.C.
(
The Office Action, at page 4, states that claims 1, 4, 12, 13, 21, 31, 39, and 40 are rejected under pre-AIA 35 USC § 103(a) over EP 1574486 A2 to Cho in view of WO 2005/103106 Al to Kumacheva and WO 2005/010145 to A2 to Dressman. At page 10, claims 32-34 are rejected over Cho, Kumacheva, and Dressman and further in view of Dorfman, 2005, Analytic Chem 77:3700-3704.
Applicant submits that the claims are not obvious.
When a patent claim includes a limitation that is not found in the prior art, then that prior art does not support a conclusion of obviousness. Arendi S.A.R.L. v. Apple, Inc., 832 F.3d 1355, 1366-67 (Fed. Cir. 2016).
Independent claim 1 recites breaking droplets by introducing a droplet-destabilizing second fluid comprising a destabilizing surfactant to thereby recover an aqueous sample fluid including an extension product from the plurality of droplets.
Independent claim 46 recites recovering an aqueous sample fluid including an extension product from a plurality of droplets by introducing a droplet-destabilizing second fluid comprising a perfluorinated alcohol.
As stated at page 14 of the Office Action, the prior art does not teach or suggest a method that includes introducing a droplet-destabilizing second fluid that includes a destabilizing surfactant or a perfluorinated alcohol allowing aqueous phase components from the emulsions to be recovered in a minimum number of steps and in a gentle manner.
Applicant respectfully submits that the independent claims are not obvious. Because the independent claims are not obvious, none of the claims are obvious. Applicant submits that the rejections under @ 103 are overcome.
Conclusion


CITED REFERENCE SUMMARIES: Ref WO2005103106A1: Claims:
A process for producing polymer particles of predetermined size and/or shape, and /or morphology, comprising the steps of: a) injecting a first fluid comprising a constituent which can harden into a microfluidic channel; b) injecting at least a second fluid into the microfluidic channel for causing the first fluid to forms into fluidic droplets within the at least second fluid causing the fluidic droplets to flow through the microfluidic channel, the microfluidic channel being sufficiently long so that the fluidic droplets harden into particles of predetermined size and/or shape while flowing through the channel; and c) collecting the hardened particles of predetermined size and/or shape from the microfluidic, channel.
The process according to claim 1 , wherein the fluidic droplets harden by any one of ionic crosslinking, hydrogen bonding, chelation, complexation and combinations thereof.
The process according to claim 1 , wherein the first fluid comprises a polymerizable constituent, and wherein said fluidic droplets harden by polymerizing into polymer particles.
The process according to claims 1 , 2 or 3 wherein the microfluidic channel is constructed and configured to produce fluidic droplets having a pre-selected size and or shape.
The process according to claims 1 , 2, 3 or 4 wherein the microfluidic channel has a cross-sectional shape being any one of circular, oval, triangular, irregular, square, and rectangular.
The process according to claims 1 , 2, 3, 4 or 5 wherein the microfluidic channel has an aspect ratio (length to average cross sectional dimension) of at least about 10:1.
The process according to claims 1 , 2, 3, 4 or 6 wherein the first and second fluids are each injected with a controlled flow rate into the microfluidic channel.
The process according to claims 1 , 2, 3, 4, 5, 6 or 7 wherein a diameter of the fluidic droplets formed in step b) is controlled by controlling flow rates and flow rate ratios of the first and second fluids, controlling dimensions of said microfluidic channel, properties of said first and second fluids including viscosity and interfacial tension between the first and second fluids, controlling properties of the polymerizable constituent, and wherein said microfluidic channel includes a portion constructed with pre-selected cross-sectional shape and dimensions to give a desired shape to said fluidic droplets upon transit of said fluidic droplets through said portion of said fluidic channel.
The process according to any one of claims 1 to 8 wherein said fluidic droplets have a diameter d, and wherein said portion of the microfluidic channel is constructed with pre-selected cross-sectional shape and dimensions selected so that d is larger than at least one of the dimensions of the microfluidic channel so that fluidic droplets with non-spherical shapes are formed upon transit through said portion of the microfluidic channel with preselected cross-sectional shape and dimensions so that said collected polymer particles have a non-spherical shape.
The process according to claim 9 wherein an aspect ratio for said non- spherical droplets is varied by changing a ratio between fluidic droplet volume and said dimensions of the microfluidic channel.
The process according to any one of claims 1 to 8 wherein said fluidic droplets have a diameter d, and wherein said portion of the microfluidic channel has a width w and a height h, and wherein when said portion of the  microfluidic channel is constructed so that w > d and h > d, and a flow rate of the fluidic droplets through the microfluidic channel is selected to be slow enough so that the fluidic droplets maintain a spherical shape so that said collected polymer particles have a spherical shape.
The process according to ' any one of claims 1 to 8 wherein said fluidic droplets have a diameter d, and wherein said portion of the microfluidic channel has a width wand a height h, and wherein when said portion of the microfluidic channel is constructed so that w > d and h > d, and a flow rate of the fluidic droplets through the microfluidic channel is selected to be high enough so that the fluidic droplets acquire an ellipsoidal shape so that said collected polymer particles have an ellipsoidal shape.
The process according to any one of claims 1 to 8 wherein said fluidic droplets have a diameter d, and wherein said portion of the microfluidic channel has a width w and a height h, and wherein when said portion of the microfluidic channel is constructed so that w < d and h > d the droplets acquire a discoid shape so that said collected polymer particles have a discoid shape shape.
The process according to any one of claims 1 to 8 wherein said fluidic droplets have a diameter d, and wherein said portion of the microfluidic channel has a width w and a height h, and wherein when said portion of the microfluidic channel is constructed so that w < d, h < d the droplets acquire a rod shape so that said collected polymer particles have a rod shape.
The process according to any one of claims 1 to 8, and 11 to 14 wherein said at least a second fluid includes at least two immiscible fluids, and wherein said two immiscible fluids and said first fluid are injected into adjacent microfluidic channel inlets, and wherein one of said two immiscible fluids envelops the first fluid mixed with the other immiscible fluid thereby forming a coaxial stream which during passage through said microfluidic channel breaks up into segments, and wherein said segments form spherical fluidic cores enveloped by a shell which form core/shell particles.
The process according to claim 15 including adjusting flow rates of each of said first fluid and said at least two immiscible fluids to selectively control a size of the cores, a thickness of shells, and a size of core-shell particles, and a number of cores per particle.
The process according to claim 16 wherein there are at least five adjacent microfluidic channel inlets including a middle microfluidic channel inlet, two inner microfluidic channels with said middle microfluidic channel inlet being located between said two inner microfluidic channel inlets, and two outer microfluidic channel inlets with one outer outer microfluidic channel inlet being located adjacent to one of the inner microfluidic channel inlets and the other outer microfluidic channel inlet being located adjacent to the other inner microfluidic channel inlet, and wherein said first fluid is injected into said middle microfluidic channel inlet, one of the at least two immiscible fluids being injected into the two inner microfluidic channel inlets and the other of the at least two immiscible fluids being injected into the two outer microfluidic channel inlets.
The process according to claim 17 wherein the immiscible fluid injected into the two inner microfluidic channel inlets is an oil, and wherein the immiscible fluid injected into the two outer microfluidic channel inlets is an aqueous fluid.
The process according to claim 17 wherein said coaxial stream becomes unstable to perturbations with wavelengths larger than a circumference of said coaxial stream which causes the coaxial stream to break up into segments,
The process according to claim 16, 17, 18 or 19 including controlling a number of cores per fluid droplet by adjusting relative flow rates of the first fluid and the at least two fluids, and by varying interfacial capillary wavelengths Î» m and Î» 0 , where Î» m is an interfacial capillary wavelength of said monomer, and Î» 0 is an interfacial capillary wavelength of said second fluid.
The process according to any one of claims 1 to 20 wherein the constituent which can harden is a polymerizable constituent including a mixture of two or more monomers, oligomers, liquid polymers or combinations thereof such that said polymer particles are copolymer particles.
The process according to any one of claims 1 to 20 wherein the constituent which can harden is a polymerizable constituent including a monomer or oligomer or liquid polymer.
The method according to any one of claims 1 to 22 wherein the second fluid is immiscible with the first fluid so that the first fluid forms fluidic droplets.
The process according any one of claims 1 to 23 wherein the collected hardened particles of pre-selected size and/or shape and /or morphology are formed into a product.
The process according to any one of claims 1 to 24 wherein said microfluidic channel is formed in a microreactor.
The process according to claim 25 wherein said microreactor comprises a hydrophobic material, and wherein said first and second fluids exhibit different degrees of hydrophobicity, and wherein the less hydrophobic of the first and second fluids forms the fluidic droplets in the other.
The process according to claim 25 wherein said microreactor comprises a hydrophilic material, and wherein said first and second fluids exhibit different degrees of hydrophilicity and wherein the less hydrophilic of the first and second fluids forms the fluidic droplets in the other.
The process according to claim 25 wherein said microreactor comprises a moderately hydrophobic material.
The process according to any one of claims 1 to 28, wherein said first  fluid comprises one or more of organic dyes, chromophores, nonlinear optical compounds, fluorescent dyes, inorganic chemicals, inorganic particles, inorganic pigments, fluorescent inorganic particles, semiconductor nanoparticles (quantum dots), inorganic particles having pre-selected magnetic properties, inorganic particles having pre-selected anti-magnetic properties, inorganic particles having pre-selected electrically conductive and/or semiconducting properties, carbon nanotubes, liquids such as liquid crystal, which are incorporated into said hardened particles, wherein said hardened particles are composite hardened particles.
The process according to any one of claims 1 to 29 wherein said hardened particles of predetermined size and/or shape and /or morphology are monodisperse.
The process according to claim 30 wherein no more- than about 5% of the hardened particles have a dimension greater than about 5% of the average dimension.
The process according to claim 30 wherein no more than about 3% of the hardened particles have a dimension greater than about 10% of the average dimension.
The process according to any one of claims 1 to 32, wherein the hardened particles have an average dimension of less than about 100 micrometers.
The process according to any one of claims 1 to 32, wherein the hardened particles have a shape selected from the group consisting of spherical, disk, rod, oval, ellipsoidal, plates, truncated spheres, hemispheres and bowls.
The process according to any one of claims 1 to 34, wherein the hardened particles are hollow.
The process according to any one of claims 1 to 34, wherein the hardened particles have a core/shell structure with a controlled number of cores.
The process according to any one of claims 1 to 36, wherein the first fluid comprises a polymerizable constituent including carbon or carbon-wall nanotubes so that the hardened particles are polymer particles having carbon or carbon-wall nanotubes incorporated therein.
The process according to any one of claims 1 to 37, including injecting an unpolymerizable liquid into the microfluidic channel along with the first and second fluids wherein some of said unpolymerizable liquid is incorporated into the polymer particles.
The process according to any one of claims 1 to 37, wherein said first fluid comprises a polymerizable constituent which includes an unpolymerizable liquid wherein some of said unpolymerizable liquid for producing polymer particles having unpolymerizable liquids incorporated into the polymer particles.
The process according to claim 38 or 39, wherein the unpolymerizable liquid is any one of a liquid crystal, a porogen, a dispersion of inorganic or organic particles.
The process according to any one of claims 1 , including injecting an unpolymerizable fluid into the microfluidic channel along with the first and second fluids wherein said hardened particles has unpolymerizable fluid incorporated into the polymer particles for producing porous polymer particles.
The process according to any one of claims 1 to 41 , wherein said first fluid includes two or more monomers, and wherein said polymer particles include a core formed by polymerization of one of said two ore more polymers and a shell enveloping said core formed by polymerization of at least one of the other of said two ore more polymers.
The process according to claim 25, wherein said first fluid includes two or more monomers includes two or more monomers, and wherein said polymer particles include one or more polymer cores formed by polymerization of one or more of said polymers, and a shell enveloping said core formed by polymerization of at least one of the other of said two ore more polymers.
The process according to any one of claims 1 to 43, wherein said first fluid comprises a biocompatible product including one or more of starch, derivatives, polymers containing 3-hydroxybutyrate and its derivatives, polymers containing 3-hydroxyvalerate and its derivatives, protein, nucleic acids (DNA, RNA), amino acid (peptide), liposomes, agaroze and its derivatives, chitosane and it's derivatives, alginate and its derivatiives, pectin and its derivatives, cellulose derivatives, drugs and their derivatives which are incorporated into said hardened particles, wherein said hardened particles are composite particles.
The process according to any one of claims 1 to 43 including exposing said fluidic droplets in said microfluidic channel to a polymerizing agent for polymerizing said fluidic droplets.
The process according to claim 45 wherein said polymerizing agent is one of heat, ultra-violet light (UV), plasma, irradiation.
The process according to any one of claims 1 to 46 including exposing said fluidic droplets in said microfluidic channel to an external field including any one or more of a magnetic field, an electric field, light or other form of radiation.
The process according to any one of claims 1 to 47 wherein said first fluid includes a gel precursor material that hardens upon chemical or physical crosslinking so that upon hardening the fluidic droplets gel to form microgel particles.
The process according to any one of claims 1 to 48 wherein said first fluid includes biological cells so that said hardened particles contain biological cells.
The process according to any one of claims 1 to 49 wherein said first fluid includes two or more monomeric, polymeric or oligomeric constituents, and wherein the microfluidic droplets harden in a stepwise manner, including exposing said microfluidic droplets to at least polymerizing agents at different positions along said microfuidic channel in a pre-selected sequence to sequentially polymerize the different constituents at different times.
The process according to any one of claims 1 to 50 wherein production of hardened particles is done in a continuous throughput process.
The process according to anyone of claims 15 to 20 wherein the cores are solid.
The process according to anyone of claims 15 to 20 wherein the cores are liquids encapsulated by said shell.
The process according to anyone of claims 1 to 53 wherein the step c) of collecting the hardened particles of predetermined size and/or shape from the microfluidic channel includes producing a lattice structure with said hardened particles.
The process according to any one of claims 1 to 43 wherein said hardened particles include two or more monodisperse populations of hardened particles of predetermined size and/or shape and/or morphology, and wherein each population has a size different from the other populations.
The process according to claim 55 wherein the step c) of collecting the two or more monodisperse populations of hardened particles includes producing a lattice structure with said two or more monodisperse populations of hardened particles.
The process according to any one of claims 1 to 33, wherein the hardened particles are porous.
The process according to any one of claims 1 to 57 wherein said first fluid includes a gel material, and including controlling flow conditions of said first and second fluids in said microfluidic channel that said first fluid is sheared by the second fluid such that it undergoes shear thinning, liquifies and brakes in droplets which harden and form microgel particles in said microfluidic channel.
The process according to any one of claims 1 to 58 wherein said , microfluidic channel is formed in a microfluidic reactor made of a polyurethane material, and wherein said polyurethane material is synthesized from a polyol or a combination of polyols and polyisocyanate or a combination of polyisocyanates using organometallic or/and amine-containing catalysts.
An apparatus for producing polymer particles with pre-determined sizes and or shapes, comprising: a microreactor having an input end including one or more fluid inlets inputs and a microfluidic channel, said microfluidic channel being sufficiently long so that fluidic droplets located in the microfluidic channel have a long enough residence time to polymerize within the microfluidic channel; and the microreactor being made of a suitable material such that upon injecting a fluid comprising a polymerizable constituent into the microreactor the fluid forms into droplets within the microfluidic channel.
The apparatus according to claim 60, wherein the microreactor is produced by soft lithography.
The apparatus according to claim 60 or 61 wherein the suitable material is a hydrophobic material, and wherein a hydrophilic polymerizable constituent is injected into the apparatus which forms droplets which are polymerized.
The apparatus according to claim 60, 61 or 62 wherein the suitable material is a hydrophobic material, and wherein a hydrophilic polymerizable constituent is injected into the apparatus which forms droplets which are polymerized.
The apparatus according to claim 63 wherein the hydrophobic material is one of polysiloxane and polyurethane.
The apparatus according to claim 64 where said polyurethane material is synthesized from a polyol or a combination of polyols and polyisocyanate or a combination of polyisocyanates using organometallic or/and amine- containing catalysts.
A polyurethane material synthesized from a polyol or a combination of polyols and polyisocyanate or a combination of polyisocyanates using organometallic or/and amine-containing catalysts.
Ref WO2005010145A2: Claims:
1. A composition comprising a plurality of beads, wherein each of said plurality of beads comprises a plurality of bound polynucleotides, wherein the polynucleotides in the composition are heterogeneous, and wherein on at least 1 % of said beads the plurality of bound polynucleotides is homogeneous.
2. The composition of claim 1 wherein on at least 5 % of said beads the plurality of bound polynucleotides is homogeneous.
3. The composition of claim 1 wherein on at least 10 % of said beads the plurality of bound polynucleotides is homogeneous.
4. The composition of claim 1 wherein on at least 50 % of said beads the plurality of bound polynucleotides is homogeneous.
5. The composition of claim 1 wherein the plurality of bound polynucleotides is greater than 100.
6. The composition of claim 1 which is a liquid.
7. The composition of claim 6 which comprises agarose.
8. The composition of claim 1 wherein the polynucleotides in the composition differ by a single nucleotide polymorphism (SNP).
9. The composition of claim 1 wherein the polynucleotides in the composition differ in the presence or absence of a mutation.
10. The composition of claim 1 wherein the polynucleotides in the composition differ in the presence or absence of an insertion.
11. The composition of claim 1 wherein the polynucleotides in the composit
[Truncated to input limit]