Cell membrane for nanopore sequencing
A nanopore sequencing, cell membrane technology, applied in the field of sequencing
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Embodiment 1
[0054] Embodiment 1 block copolymer is not cross-linked film-forming
[0055] 1. Dissolve the unmodified tri-block copolymer (6PMOXA-33PDMS-6PMOXA) in chloroform, then dry it under argon atmosphere, and dissolve the dried copolymer in n-decane or silicone oil , to obtain a lysate with a concentration of 10 mg / mL.
[0056]
[0057] 2. Build a nanopore test device, use Axopatch 200B Patch Clam (Axon Company) patch clamp and data acquisition card 1550A (Axon Company) to build a signal acquisition platform, use a groove with a 150um hole, and place the holes on both sides of the groove for the experiment. 1 mL of buffer solution (1M KCl, 10 mM tris, 1 mM EDTA, pH 8.0) was added to the pools, and the patch clamp systems were connected via Ag / AgCl respectively.
[0058] 3. Inject 2μl of 10mg / ml n-decane solution or silicone oil solution of triblock copolymer with double bonds, and form a polymer film by bubbling around the hole. At the same time, the electrode at the cis end is ...
Embodiment 2
[0062] Embodiment 2 block copolymer self-crosslinking
[0063] 1. Synthesize a tri-block copolymer (6PMOXA-33PDMS-6PMOXA) with a double bond (methacrylate) at the end group, dry it under an argon atmosphere, and dissolve the dried copolymer in n-decane Or in silicone oil, a dissolved product with a concentration of 10 mg / mL was obtained.
[0064]
[0065] 2. Build a nanopore test device, use Axopatch 200B Patch Clam (Axon Company) patch clamp and data acquisition card 1550A (Axon Company) to build a signal acquisition platform, use a groove with a 150um hole, and place the holes on both sides of the groove for the experiment. Add 1mL of buffer solution (1M KCl, 10mM tris, 1mM EDTA, pH 8.0, containing 0.5% photoinitiator, such as Irgacure 2959) to each pool, and connect the patch clamp system through Ag / AgCl respectively.
[0066] 3. Inject 2μl of 10mg / ml n-decane solution or silicone oil solution of triblock copolymer with double bonds, and form a polymer film by bubbling ...
Embodiment 3
[0071] Example 3 Block copolymer and hydrogel crosslinking (crosslinking after protein insertion first)
[0072] 1. Synthesize a triblock copolymer (6PMOXA-33PDMS-6PMOXA) with methacrylate at both ends, dry it in an argon environment, and dissolve the dried copolymer in n-decane or silicone oil In, the lysate with a concentration of 10 mg / mL was obtained.
[0073] 2. Build a nanopore test device, use Axopatch 200B Patch Clam (Axon Company) patch clamp and data acquisition card 1550A (Axon Company) to build a signal acquisition platform, use a groove with a 100 μm hole, and insert it into the fluid tank for the experiment Add 1 mL of buffer containing 10% w / v polyethylene glycol-dimethacrylic acid monomer (1M KCl, 10mM tris, 1mM EDTA, pH 8.0, containing 0.5% photoinitiator Irgacure), and pass through Ag / AgCl connection to the patch clamp system. 3. Inject 2μl of 10mg / ml n-decane solution or silicone oil solution of triblock copolymer with double bonds, and form a polymer fil...
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