{"id":38402,"date":"2023-02-18T15:55:48","date_gmt":"2023-02-18T07:55:48","guid":{"rendered":"http:\/\/biopioneer.com.tw\/?p=38402"},"modified":"2023-06-11T14:02:34","modified_gmt":"2023-06-11T06:02:34","slug":"38402","status":"publish","type":"post","link":"http:\/\/biopioneer.com.tw\/?p=38402","title":{"rendered":"\u907a\u50b3\u5de5\u7a0bDNA\u5206\u5b50\u6280\u8853 | \u805a\u5408\u9176\u93c8\u53cd\u61c9 (Polymerase Chain Reaction, PCR)\/DNA\u51dd\u81a0\u96fb\u6cf3"},"content":{"rendered":"

\u3010\u907a\u50b3\u5de5\u7a0bDNA\u5206\u5b50\u6280\u8853<\/a>\u3011<\/strong><\/span><\/p>\n

\u805a\u5408\u9176\u93c8\u53cd\u61c9 (Polymerase Chain Reaction, PCR)\/DNA\u51dd\u81a0\u96fb\u6cf3<\/span><\/strong><\/p>\n

PCR\u53cd\u61c9->\u74ca\u8102\u81a0\u9ad4\u7c89\u672b<\/a><\/span>\u914d\u88fd->DNA\u51dd\u81a0\u96fb\u6cf3\uff08agarose gel electrophoresis\uff09->\u6838\u9178\u67d3<\/a>\u8272<\/span>-><\/span><\/span><\/strong>\u5728\u96fb\u6cf3\u5b8c\u6210\u5f8c\u85c9\u7531\u85cd\u5149\u89c0\u5bdf\u7bb1<\/a><\/span>\u6fc0\u767c<\/strong>\u770b\u5230\u4e0d\u540c\u5927\u5c0f\u7684DNA\u7247\u6bb5\u3002<\/span><\/p>\n

\"PCR\u53cd\u61c9\"<\/p>\n

\u805a\u5408\u9176\u93c8\u53cd\u61c9 (Polymerase Chain Reaction, PCR)<\/span> <\/strong><\/span>\u662f\u4e00\u7a2e\u5206\u5b50\u6280\u8853(molecular technology)\uff0c\u7531\u8afe\u8c9d\u723e\u734e\u7372\u5f97\u8005 Kary Mullis \u5728 1980 \u5e74\u4ee3\u958b\u767c\uff0c\u53ef\u4ee5\u9ad4\u5916(in vivo) \u5feb\u901f\u4e14\u5ec9\u50f9\u5730\u64f4\u589e DNA \u7247\u6bb5(DNA fragments)(1)\u3002\u5b83\u5df2\u6210\u70ba\u907a\u50b3(\u00a0genetic )\u548c\u5206\u5b50\u7814\u7a76(molecular research)\u7684\u57fa\u672c\u5de5\u5177\uff0c\u56e0\u70ba DNA \u5be6\u9a57(DNA experimentation)\u901a\u5e38\u9700\u8981\u5927\u91cf\u7684\u76ee\u6a19 DNA (2)(3)\u3002\u4e00\u4e9b\u4f9d\u8cf4 PCR \u6280\u8853(PCR technology)\u7684\u61c9\u7528\u5305\u62ec DNA \u6e2c\u5e8f(DNA sequencing)\uff08\u4f8b\u5982\u4eba\u985e\u57fa\u56e0\u7d44\u8a08\u5283,Human Genome Project\uff09\u3001DNA \u6307\u7d0b\u8b58\u5225(DNA fingerprinting)\u3001\u6cd5\u91ab\u5b78(forensics)\u3001\u7d30\u83cc(bacteria)\u6216\u75c5\u6bd2(\u00a0viruses)\u6aa2\u6e2c\uff08\u5c24\u5176\u662f AIDS\uff09\u548c\u907a\u50b3\u6027\u75be\u75c5(hereditary disease)\u7684\u8a3a\u65b7 (4)(5)\u3002\u7531\u65bc\u80fd\u5920\u5f9e\u5c11\u91cf\u6838\u9178(nucleic acid)\u751f\u6210\u5927\u91cf DNA\uff0cPCR \u662f\u4e00\u7a2e\u975e\u5e38\u6709\u6548\u7684 DNA \u64f4\u589e\u65b9\u6cd5(amplify DNA )\uff0c\u56e0\u6b64\u6709\u6642\u88ab\u7a31\u70ba\u201c\u5206\u5b50\u5fa9\u5370(molecular photocopying)\u201d(2)(3)(6)\u3002<\/p>\n

\u70ba\u4e86\u6aa2\u67e5\u662f\u5426\u64f4\u589e\u4e86\u6b63\u78ba\u7684\u76ee\u6a19 DNA \u7247\u6bb5\uff0c\u51dd\u81a0\u96fb\u6cf3(gel electrophoresis)\u662f\u6aa2\u67e5\u64f4\u589e\u7522\u7269(amplified products)\u5206\u5b50\u5927\u5c0f\uff08\u4ee5 bp \u70ba\u55ae\u4f4d\uff09\u7684\u5feb\u901f\u65b9\u6cd5\u3002\u6d41\u7a0b\u5982\u4e0b\uff1aPCR\u53cd\u61c9\u7522\u7269\uff0c\u7d93\u5fae\u91cf\u5206\u6ce8\u5668\u53d6\u5c11\u91cfLoading\u81f3\u914d\u88fd\u597d\u7684\u74ca\u8102\u81a0\u9ad4<\/a><\/span>\uff0c \u653e\u7f6e<\/strong>\u6c34\u5e73\u96fb\u6cf3\u69fd<\/a><\/span>\uff0c<\/strong>\u7d93\u51dd\u81a0\u96fb\u6cf3(gel electrophoresis)\u5f8c\uff0c<\/strong>\u96fb\u6cf3\u5b8c\u7684\u74ca\u8102\u81a0\u9ad4<\/a><\/span>, \u7d50\u5408DNA\u6838\u9178\u67d3\u5291<\/a><\/span>\uff0c<\/span><\/span><\/span><\/strong>\u5728\u85cd\u5149\u89c0\u5bdf\u7bb1<\/a><\/strong><\/span>\u6fc0\u767c\u4e0b\uff0c\u89c0\u5bdf\u4e0d\u540c\u5927\u5c0f\u7684DNA\u7247\u6bb5\u3002\u7d93\u7531\u64f4\u589e\u7684 PCR \u7522\u7269\u7684\u5927\u5c0f\u662f\u901a\u904e\u5c07\u5176\u8207DNA Ladder<\/a>\u00a0\u9032\u884c\u6bd4\u8f03\u4f86\u4f30\u8a08\u7684\uff0cDNA Ladder<\/a>\u662f\u4e00\u7a2e\u5206\u5b50\u91cf\u6a19\u8a18\uff0c\u5176\u4e2d\u5305\u542b\u5df2\u77e5\u5927\u5c0f\u7684 DNA \u7247\u6bb5\u3002<\/p>\n

\"\"<\/p>\n

\u8a31\u591a\u56e0\u7d20\u6703\u5e72\u64fe PCR \u53cd\u61c9\u3002\u6709\u4e9b\u6613\u65bc\u512a\u5316\uff08\u4f8b\u5982\u71b1\u5faa\u74b0\u689d\u4ef6\u548c Mg\u00a02+\u6fc3\u5ea6\uff09\uff0c\u800c\u53e6\u4e00\u4e9b\u5247\u64cd\u4f5c\u8d77\u4f86\u6bd4\u8f03\u68d8\u624b\uff0c\u9700\u8981\u7a4d\u7d2f\u7d93\u9a57\u624d\u80fd\u89e3\u6c7a\uff08\u4f8b\u5982\u5f15\u7269\u8a2d\u8a08\u548c PCR \u7de9\u885d\u6db2\u6210\u5206\/\u6dfb\u52a0\u5291\uff09\u3002\u4e00\u822c\u4f86\u8aaa\uff0cPCR \u4e2d\u7684\u91cd\u8981\u56e0\u7d20\u5305\u62ec DNA \u6a21\u677f\u3001DNA \u805a\u5408\u9176\u3001\u5f15\u7269\u8a2d\u8a08\u3001\u7de9\u885d\u6db2\u6210\u5206\u3001\u6dfb\u52a0\u5291\u548c\u6291\u88fd\u5291\u4ee5\u53ca\u71b1\u5faa\u74b0\u689d\u4ef6 (6)\u3002\u6216\u9078\u64c72XPCR\u6df7\u5408\u6db2<\/a>\u9032\u884c PCR \u53cd\u61c9\uff0c2X PCR \u6df7\u5408\u6db2 (2X PCR SuperMix, MB200-P100 )\u5177\u6700\u4f73\u4e4bMg ++\u3001dNTP\u548c\u91cd\u7d44Taq DNA\u805a\u5408\u9176\u7684\u6df7\u5408\u7269\u3002\u50c5\u9700\u52a0\u5165 template DNA\u3001primer \u6df7\u5408\u6db2\u5373\u53ef\u958b\u59cb\u9032\u884c PCR \u53cd\u61c9\u3002<\/p>\n

\u74ca\u8102\u81a0\u9ad4\u7684\u88fd\u5099\uff1a\u53d6\u9069\u91cf\u74ca\u8102\u81a0\u9ad4\u7c89\u672b<\/a>\u653e\u5165\u7de9\u885d\u6db2\uff08\u5e38\u7528\u7684\u662fTBE\u7de9\u885d\u6db2\uff09\u4e2d\uff0c\u4ee5\u5fae\u6ce2\u52a0\u71b1\u81f3\u5b8c\u5168\u6eb6\u89e3\uff0c\u6df7\u5408\u5747\u52fb\u5f8c\uff0c\u51b7\u537b\u81f3\u7d0450\u2103\uff0c\u5012\u5165\u63d2\u6709\u9f52\u72c0\u6a21\u578b\u4e4b\u9444\u6a21\u4e2d\uff0c\u975c\u7f6e\u7b49\u5f85\u81a0\u9ad4\u51dd\u56fa\u5f8c\uff0c\u518d\u53d6\u4e0b\u9f52\u72c0\u6a21\u578b\u3002\u53d6\u51fa\u5df2\u5b8c\u5168\u51dd\u56fa\u7684\u81a0\u9ad4\uff0c\u7f6e\u65bc\u96fb\u6cf3\u69fd\u5167\uff0c\u4e26\u52a0\u5165\u7de9\u885d\u6db2\u81f3\u8986\u84cb\u81a0\u9ad4\u3002<\/p>\n

\"PCR\u8a66\u5291_\u5de5\u4f5c\u5340\u57df<\/a><\/span><\/strong><\/p>\n

PCR\u6df7\u5408\u6db2<\/a>\u00a0| 100bp-10kb DNA Ladder\u00a0<\/a>\u00a0|\u00a0\u74ca\u8102\u81a0\u9ad4\u7c89Agarose<\/a>\u00a0|\u00a0\u00a0DNA\u6838\u9178\u67d3\u5291<\/a><\/span><\/span><\/strong><\/p>\n

\"BIO-HELIX\"\"BIO-HELIX<\/a>\"BIO-HELIX<\/a>\"BIO-HELIX<\/p>\n

 <\/p>\n

\u805a\u5408\u9176\u934a\u5f0f\u53cd\u61c9 (PCR) \u662f Kary Mullis \u5728 1980 \u5e74\u4ee3\u958b\u767c\u7684\u9769\u547d\u6027\u65b9\u6cd5 [1]\uff0c\u70ba\u5206\u5b50\u751f\u7269\u5b78\u4e2d\u6700\u5f37\u5927\u7684\u6280\u8853\u4e4b\u4e00\u3002\u5e8f\u5217\u7279\u7570\u6027\u5f15\u5b50\u3001\u71b1\u7a69\u5b9a DNA \u805a\u5408\u9176\u548c\u71b1\u5faa\u74b0\u5c07 DNA \u6216 cDNA \u6a21\u677f\u4e2d\u7684\u7279\u5b9a\u5e8f\u5217\u5f9e\u5c11\u91cf\u64f4\u589e\u5230\u6578\u5343\u5230\u767e\u842c\u500d\u3002 20-40 \u6b21\u91cd\u8907\u7684\u52a0\u71b1\u548c\u51b7\u537b\u5faa\u74b0\uff08\u7a31\u70ba\u71b1\u5faa\u74b0\/thermal cycling\uff09\uff0c\u901a\u904e\u9176\u4fc3\u53cd\u61c9\u4fc3\u9032 DNA \u8907\u88fd(DNA replication)\u3002\u76ee\u6a19\u5e8f\u5217\u7684\u6578\u91cf\u96a8\u8457\u6bcf\u500b\u5faa\u74b0\u52a0\u500d\uff0c\u6307\u6578\u64f4\u589e\u300240 \u6b21\u91cd\u8907\u7684 PCR \u71b1\u5faa\u74b0\u5c07\u5f9e\u6a21\u677f\u7684\u55ae\u500b\u62f7\u8c9d\u751f\u6210 1,099,511,627,776 \u500b DNA \u62f7\u8c9d\u3002<\/p>\n

In a nutshell, PCR is essentially a series of 20-40 repeated cycles of heating and cooling (called thermal cycling) that facilitate DNA replication through enzymatic reactions. The amount of target sequence doubles with each cycle which leads to an exponential amplification represented by 2(# of cycles)<\/sup>. Therefore, a PCR thermal cycling of 40 repeats will generate 1,099,511,627,776 copies of DNA from a single copy of the template. The function and purpose of each step in a PCR Reaction are discussed below\u00a0(Figure 1)<\/b>:<\/p>\n

\"\"<\/p>\n

Figure 1<\/b>\u00a0\u2013 How Polymerase Chain Reaction works: 1) Denaturation: the reaction is heated to 94\u201398 \u00b0C for 20\u201330 seconds to break the hydrogen bonds between the strands. 2) Annealing: the reaction temperature is lowered to 50-65 \u00b0C for 20-40 seconds to allow primers to anneal to the template strands. 3) Elongation: the temperature is increased (the optimal temperature is dependent on the DNA Polymerase used e.g. 72-78 \u00b0C for Taq Polymerase) to allow for the addition of dNTPs. The amount of target sequence doubles with each thermal cycle which leads to an exponential amplification represented by 2(# of cycles)<\/sup><\/p>\n

\u8d77\u59cb\u53cd\u61c9( Initialization)<\/span><\/strong><\/span><\/p>\n

\u5c07\u53cd\u61c9\u52a0\u71b1\u81f3 94\u201396\u00b0C\uff08\u5982\u679c\u4f7f\u7528\u6975\u8010\u71b1\u7684 DNA \u805a\u5408\u9176\uff0c\u5247\u52a0\u71b1\u81f3 98\u00b0C\uff092-10 \u5206\u9418\u3002\u8a72\u6b65\u9a5f\u6fc0\u6d3b\u53cd\u61c9\u4e2d\u7684 DNA \u805a\u5408\u9176\u548c\u8a66\u5291\uff0c\u4e26\u4f7f\u6df7\u5408\u7269\u4e2d\u7684\u5176\u4ed6\u6c61\u67d3\u7269\uff08\u5982\u679c\u6709\uff09\u8b8a\u6027\u3002\u5728\u83cc\u843d\u7be9\u9078\u7b49\u61c9\u7528\u4e2d\uff0c\u53ef\u76f4\u63a5\u4f7f\u7528\u5c11\u91cf\u7d30\u80de\u4f5c\u70ba\u6a21\u677f\u3002\u6b64\u521d\u59cb\u5316\u6b65\u9a5f\u5c07\u88c2\u89e3\u7d30\u80de\u4ee5\u91cb\u653e DNA \u4e26\u4f7f\u5176\u4ed6\u7d30\u80de\u86cb\u767d\u8cea\uff08\u5305\u62ec DNase\uff09\u8b8a\u6027\u3002\u5982\u679c\u5728 PCR \u671f\u9593\u4f7f\u7528\u5316\u5b78\u54c1\u6216\u6297\u9ad4\uff08\u4f8b\u5982 BlasTaq\u2122 HotStart DNA \u805a\u5408\u9176\uff09\uff0c\u5247\u5fc5\u9808\u57f7\u884c\u6b64\u6b65\u9a5f\u4ee5\u71b1\u6fc0\u6d3b\u9176\u3002<\/p>\n

\u8b8a\u6027(Denaturation)<\/span><\/strong><\/span><\/p>\n

\u5c07\u53cd\u61c9\u52a0\u71b1\u81f3 94\u201398\u00b0C \u4e26\u4fdd\u6301 20\u201330 \u79d2\u3002\u9019\u662f\u71b1\u5faa\u74b0(thermal cycling)\u7684\u7b2c\u4e00\u6b65\u3002\u96d9\u93c8 DNA(Double-stranded DNA) \u5728\u6b64\u6b65\u9a5f\u4e2d\u8b8a\u6027\uff0c\u56e0\u70ba\u96d9\u93c8\u4e4b\u9593\u7684\u6c2b\u9375(hydrogen bonds)\u56e0\u9ad8\u6eab\u800c\u65b7\u88c2\uff0c\u9019\u4e00\u904e\u7a0b\u4e5f\u7a31\u70ba DNA \u7194\u5316(DNA melting)\u3002
\n\u93c8\u5408(Annealing)<\/strong><\/span><\/p>\n

\u5f15\u5b50(Primers )\u662f\u53ef\u4ee5\u8207 DNA \u6a21\u677f(DNA template)\u7684\u7279\u5b9a\u5e8f\u5217(specific sequences)\u7d50\u5408\u4ee5\u6307\u5c0e DNA \u805a\u5408\u9176\u8907\u88fd\u7684\u5be1\u6838\u82f7\u9178(\u00a0oligonucleotides )\u3002\u70ba\u4f7f\u5f15\u7269(\u00a0primers )\u8207\u55ae\u93c8 DNA \u6a21\u677f\u00a0annealing\uff0c\u901a\u5e38\u5c07\u53cd\u61c9\u6eab\u5ea6\u964d\u81f3 50-65\u00b0C\uff0c\u6301\u7e8c 20-40 \u79d2\u3002Annealing\u7684\u6700\u4f73\u6eab\u5ea6\u53d6\u6c7a\u65bc\u5f15\u7269\u7684\u89e3\u93c8\u6eab\u5ea6 (T\u00a0m\u00a0)\uff1aDNA \u96d9\u93c8\u9ad4(DNA duplex)\u7684\u4e00\u534a\u89e3\u96e2\u6210\u55ae\u93c8\u7684\u6eab\u5ea6\u3002Annealing\u6eab\u5ea6\u8a2d\u7f6e\u904e\u9ad8\u5f15\u7269\u4e0d\u80fd\u8207\u6a21\u677fAnnealing\uff0cAnnealing\u6eab\u5ea6\u904e\u4f4e\u53ef\u80fd\u767c\u751f\u975e\u7279\u7570\u6027\u5f15\u7269\uff08\u5c0e\u81f4\u975e\u7279\u7570\u6027\u64f4\u589e\uff09\u3002\u56e0\u6b64\uff0c\u7406\u60f3\u7684Annealing\u6eab\u5ea6\u901a\u5e38\u6bd4 Tm \u4f4e 3-5\u00b0\u00a0C\u7684\u5f15\u7269((Primers )\uff0c\u8db3\u5920\u9ad8\u4ee5\u9032\u884c\u7279\u7570\u6027\u5f15\u5b50(Primers ) )Annealing\uff0c\u4f46\u53c8\u8db3\u5920\u4f4e\u4ee5\u5141\u8a31\u6709\u6548\u7684\u5f15\u5b50(Primers )\u3002\u53cd\u61c9\u6df7\u5408\u7269\u4e2d\u7684\u5f15\u5b50(Primers )\u6fc3\u5ea6\u901a\u5e38\u9060\u9ad8\u65bc DNA \u6a21\u677f\u7684\u6fc3\u5ea6\uff0c\u56e0\u6b64\u5f15\u5b50(Primers )-\u6a21\u677f\u96dc\u4ea4\u6bd4\u6a21\u677f\u93c8\u7684\u91cd\u65b0Annealing\u66f4\u53d7\u9752\u775e\u3002\u4e00\u65e6\u5f15\u5b50(Primers )\u8207\u6a21\u677fAnnealing\uff0cDNA \u805a\u5408\u9176\u5c31\u53ef\u4ee5\u958b\u59cb\u5c07 dNTP \u7d50\u5408\u5230\u6a21\u677f\u4e0a\u3002<\/p>\n

\u5ef6\u4f38\/\u4f38\u9577(Extension\/Elongation)<\/span><\/strong><\/span><\/p>\n

\u5728\u6b64\u6b65\u9a5f\u4e2d\uff0cDNA \u805a\u5408\u9176\u8207\u5f15\u5b50(Primers )\/\u6a21\u677f(template)\u8907\u5408\u7269\u7d50\u5408\uff0c\u4e26\u958b\u59cb\u5728\u5408\u6210\u93c8(synthesizing strand)\u4e0a\u4ee5 5′ \u5230 3′ \u7684\u65b9\u5411\u647b\u5165 dNTP\u3002\u9019\u6703\u7522\u751f\u4e00\u689d\u65b0\u5408\u6210\u7684\u8207\u6a21\u677f\u93c8(template strand)\u4e92\u88dc\u7684 DNA \u93c8\u3002\u5c0d\u65bc\u4e0d\u540c\u7684 DNA \u805a\u5408\u9176(\u00a0DNA polymerases)\uff0c\u5ef6\u4f38\/\u5ef6\u9577(extension\/elongation )\u7684\u6700\u4f73\u6eab\u5ea6\u4e0d\u540c\u3002Taq DNA \u805a\u5408\u9176(DNA polymerases)\uff08\u8010\u71b1\uff09\u7684\u7406\u60f3\u5de5\u4f5c\u6eab\u5ea6\u70ba 72-78\u00b0C\u3002\u5ef6\u4f38\u7684\u6301\u7e8c\u6642\u9593\u53d6\u6c7a\u65bc\u539f\u59cb\u6a21\u677f\uff08\u53c8\u540d\u64f4\u589e\u5b50\uff09\u7684\u9577\u5ea6\u548c DNA \u805a\u5408\u9176\u7684\u901f\u5ea6\u3002\u5178\u578b\u7684 DNA \u805a\u5408\u9176(\u00a0Typical DNA polymerases)\u5728\u5176\u6700\u4f73\u6eab\u5ea6\u7bc4\u570d\u5167\u4ee5 1-1.5kb\/min \u7684\u901f\u5ea6\u805a\u5408\u3002<\/p>\n

\u6700\u7d42\u4f38\u9577\u7387(Final Elongation)<\/span><\/strong><\/p>\n

\u5728\u6700\u5f8c\u4e00\u500b\u5ef6\u4f38\u5faa\u74b0\u4e2d\uff0c\u5c07\u53cd\u61c9\u6df7\u5408\u7269\u4fdd\u6301\u5728 72-78\u00b0C\uff08\u5927\u591a\u6578\u805a\u5408\u9176\u7684\u6700\u4f73\u5de5\u4f5c\u6eab\u5ea6\uff095-15 \u5206\u9418\u3002\u6b64\u6b65\u9a5f\u78ba\u4fdd\u4efb\u4f55\u5269\u9918\u7684\u55ae\u93c8 DNA \u5728\u6700\u5f8c\u4e00\u500b PCR \u5faa\u74b0\u5f8c\u5b8c\u5168\u5ef6\u4f38\u3002
\n\u6700\u5f8c\u4fdd\u6301(Final Hold)<\/strong><\/span><\/p>\n

\u5c07\u6eab\u5ea6\u964d\u4f4e\u81f3 4-15\u00b0C \u4e26\u6301\u7e8c\u7121\u9650\u9577\u7684\u6642\u9593\u4ee5\u77ed\u671f\u5132\u5b58\u53cd\u61c9\u6df7\u5408\u7269 (6)\u3002<\/p>\n

\"HOT\"<\/a><\/span><\/strong>Bio-Helix\u00a0 2X PCR SuperMix\u00a0PCR\u6df7\u5408\u6db2| \u8ca8\u865f\u00a0MB200-P100\u00a0( BIO-HELIX , 100\u500b\u53cd\u61c9)<\/a>\u00a025UL\/\u53cd\u61c9\u00a0<\/span><\/strong><\/span><\/p>\n

100 rxns (2 x 1.25mL)\u00a025UL\/\u53cd\u61c9\u00a0<\/span><\/strong><\/p>\n

\"BIO-HELIX\"<\/a><\/p>\n

\u7522\u54c1\u63cf\u8ff0:<\/strong>\u00a0<\/span>2X PCR \u6df7\u5408\u6db2 (2X PCR SuperMix, MB200-P100 )\u5177\u6700\u4f73\u4e4bMg ++\u3001dNTP\u548c\u91cd\u7d44Taq DNA\u805a\u5408\u9176\u7684\u6df7\u5408\u7269\u3002\u50c5\u9700\u52a0\u5165 template DNA\u3001primer \u6df7\u5408\u6db2\u5373\u53ef\u958b\u59cb\u9032\u884c PCR \u53cd\u61c9\u3002\u901a\u904e\u805a\u5408\u9176\u93c8\u53cd\u61c9\uff08polymerase chain reaction, PCR\uff09\u9ad8\u6548\u64f4\u589e\u6838\u9178\u6a21\u677f\u3002\u6bcf\u6b21\u53cd\u61c9\u9ad4\u7a4d\u70ba50ul(25ul\/\u53cd\u61c9)\uff0c\u6574\u7d44\u53ef\u63d0\u4f9b100\u500b\u53cd\u61c9\u3002\u00a02X PCR SuperMix\u975e\u5e38\u7a69\u5b9a<\/strong>\uff0c\u57284\u00b0C\u4e0b\u4fdd\u5b5812\u500b\u6708\u5f8c\uff0c\u672a\u89c0\u5bdf\u5230PCR\u6027\u80fd\u6216\u9176\u6d3b\u6027\u964d\u4f4e\u3002\u91cd\u8907\u7684\u51cd\u878d\u5faa\u74b0\u4e0d\u6703\u964d\u4f4e\u6027\u80fd\u6216\u6d3b\u6027\u3002<\/p>\n

\"\"<\/p>\n

\u5132\u5b58\u548c\u8010\u71b1\u6027\uff1a<\/strong><\/span>\u572837\u00b0C\u7684\u7a69\u5b9a\u6eab\u5ea6\u4e0b21\u5929\u3002<\/p>\n

\u8d85\u9ad8\u901f\uff1a<\/strong><\/span>\u572860\u79d2\u5167\u5b8c\u62102 kb\uff08\u64f4\u5c55\u901f\u5ea6\u70ba30b\u301c50b \/ sec\uff09\u3002<\/p>\n

\u9ad8\u9748\u654f\u5ea6\uff1a<\/strong><\/span>\u6a21\u677f\u64f4\u589e\u7684\u62f7\u8c9d\u6578\u4f4e\u3002<\/p>\n

\u5f9eLambda DNA\u64f4\u589e\u9577\u90545kb\u00a0(\u00a0Lambda DNA)\u3002<\/p>\n

\"\"<\/strong><\/p>\n

\"\"<\/p>\n

 <\/p>\n

\"HOT\"Agarose\/\u74ca\u8102\u81a0\u9ad4\u7c89\u672b\uff08\u5206\u751f\u5be6\u9a57\u7b49\u7d1a\uff09BIO-HELIX\u00a0 \u578b\u865fMB755-0500<\/a><\/span><\/strong><\/span><\/span><\/p>\n

\"BIO-HELIX<\/p>\n

Bio-Helix Agarose\u5728\u51dd\u81a0\u96fb\u6cf3\uff08agarose gel electrophoresis\uff09\u4e3b\u8981\u7528\u65bc\u5206\u96e2\u5927\u7684DNA\u6216\u8005RNA\u5206\u5b50\uff0c\u662f\u5927\u5c0f\u6838\u9178\u5206\u96e2\u4e4b\u7406\u60f3\u74ca\u8102\u81a0\u9ad4\u3002\u5177\u5206\u751f\u7b49\u7d1a, \u53ef\u63d0\u4f9b\u6bcf\u7a2e\u51dd\u81a0\u90fd\u80fd\u6e05\u6670\u5730\u5206\u8fa8DNA\uff0c\u6279\u6b21\u5230\u6279\u6b21\u5177\u4e00\u81f4\u5206\u8fa8\u7387\u3002\u4f4eEEO\u7279\u6027\u53ef\u5be6\u73feDNA\u7684\u9ad8\u96fb\u6cf3\u9077\u79fb\u7387\u3002\u5176\u5f37\u5927\u7684\u51dd\u81a0\u5f37\u5ea6\u53ef\u4ee5\u8f15\u9b06\u8655\u7406\uff0c\u6e1b\u5c11\u7834\u640d\u3002\u7522\u54c1\u6c92\u6709\u53ef\u6aa2\u6e2c\u7684DNase\u548cRNase\u6d3b\u6027\u3002(\u82e5\u9700\u7887\u72c00.5gx1 BIO-HELIX , \u8ca8\u865f#AGT002-0500)<\/p>\n

\"\u64cd\u4f5c\u65b9\u6cd5\"<\/p>\n

\"\"<\/p>\n

\"HOT\"DNA\u6838\u9178\u67d3\u5291 | Novel Green Plus (20000X) BIO-HELIX\u8ca8\u865fLD003-0500<\/a><\/span><\/strong><\/p>\n

Novel Green Plus \u662f\u4e0b\u4e00\u4ee3 DNA \u7d50\u5408\u67d3\u6599\uff0c\u53ef\u78ba\u4fdd\u9ad8\u9748\u654f\u5ea6\u548c\u8cea\u91cf\u3002\u8207\u6eb4\u5316\u4e59\u9320 (Ethidium bromide, EtBr) \u76f8\u6bd4\uff0cNovel Green Plus \u5728\u7d2b\u5916\u900f\u7167\u4e0b\u662f\u4e00\u7a2e\u53cd\u61c9\u9748\u654f\u7684 DNA \u67d3\u8272\u8a66\u5291\uff0c\u662f\u6aa2\u6e2c\u74ca\u8102\u7cd6\u51dd\u81a0(agarose gel)\u4e0a dsDNA \u6700\u9748\u654f\u7684\u67d3\u6599\u4e4b\u4e00\u3002\u67d3\u6599\u662f\u6839\u64da\u8207 PCR \u76f8\u95dc\u7684\u7279\u6027\u8a2d\u8a08\u7684\uff0c\u4f8b\u5982\u67d3\u6599\u7684 PCR \u6291\u5236\u3001\u7a69\u5b9a\u6027\u548c\u87a2\u5149\u5149\u8b5c\u3002Novel Green Plus \u70ba\u7528\u6236\u5e36\u4f86\u66f4\u53ef\u9760\u548c\u66f4\u5b89\u5168\u7684\u9ad4\u9a57\uff0c\u56e0\u70ba\u67d3\u8272\u7684\u51dd\u81a0\u53ef\u4ee5\u7528\u85cd\u5149\u900f\u7167\u5100\u53ef\u8996\u5316\uff0c\u5f9e\u800c\u907f\u514d\u4e86\u76ae\u819a\/\u773c\u775b\u7d2b\u5916\u7dda\u640d\u50b7\u7684\u98a8\u96aa\uff0c\u4e26\u6e1b\u5c11\u4e86 DNA \u4fee\u98fe\u5f15\u8d77\u7684\u526f\u4f5c\u7528\u3002<\/p>\n

\"BIO-HELIX<\/a><\/p>\n

\"HOT\"100bp-10kb DNA Ladder \u5206\u5b50\u91cf\u6a19\u6e96\u6db2\u00a0|\u00a0Bio-helix\u8ca8\u865fDM015-R500\u00a0\u00a0<\/a><\/span><\/p>\n

\u7522\u54c1\u63cf\u8ff0\uff1a<\/strong><\/span><\/p>\n

DNA\u5206\u5b50\u91cf\u6a19\u6e96\u6db2\u00a0(\u8ca8\u865f\u00a0Bio-Helix- DM015-R500\u00a0<\/span>; \u5206\u5b50\u91cf:\u00a0 \u00a0100-10K bp)\uff0c\u53ef\u5728\u9032\u884c\u74ca\u8102\u7cd6\u51dd\u81a0\u96fb\u6cf3\u6642\uff0c\u4f5c\u70baDNA\u5927\u5c0f\u7684\u5206\u5b50\u91cf\u6a19\u6e96\u4f9d\u64da\u3002DNA\u5206\u5b50\u91cf\u6a19\u6e96\u6db2\uff0c\u662fPCR\u7522\u7269\u548c\u8a31\u591a\u5c08\u6709\u8cea\u7c92\u7684\u7368\u7279\u7d44\u5408\uff0c\u7d93\u9069\u7576\u7684\u9650\u5236\u9176\u6d88\u5316\u7522\u751f13\u500b\u7247\u6bb5\u00a0(100bp\u3001250bp\u3001500bp\u3001750bp\u30011kb\u30011.5kb<\/span>\u30012kb\u30013kb<\/span>\u30014kb\u30015kb\u30016kb\u30018kb\u300110kb)<\/span>\uff0c DNA\u5305\u62ec100-10,000 base pairs\u500b\u9e7c\u57fa\u5c0d\u7684\u7247\u6bb5\u3002\u5176\u4e2d1kb\u548c3k\u5177\u8f03\u5f37\u5ea6\u4f5c\u70ba\u53c3\u8003\u9ede\u3002 \u63d0\u4f9b\u6bcf\u500b\u689d\u5e36\u4e2dDNA\u7684\u8fd1\u4f3c\u8cea\u91cf\uff080.5\u03bcg\u8f09\u8377\uff09\uff0c\u7528\u65bc\u8fd1\u4f3c\u76f8\u4f3c\u5927\u5c0f\u7684\u76f8\u5c0d\u5f37\u70c8\u6a23\u54c1\u4e2dDNA\u7684\u8cea\u91cf\u3002 \u7bc4\u570d100 – 10,000 bp\u3002\u7247\u6bb5Number of bands : 13\u3002\u5305\u88ddPackage :\u00a0\u00a0<\/strong>50\u03bcg \/ 500\u03bcl\u3002\u64cd\u4f5c\u624b\u518a\u00a0<\/a><\/strong><\/span><\/p>\n

\"\"\"Bh<\/p>\n

Note: \/ DNA Marker \/ DNA Standard
\nCategories:\u00a0DNA Marker\/Ladder<\/a><\/p>\n

\"BIO-HELIX<\/a><\/p>\n

A unique combination of PCR products and a number of proprietary plasmids digested with appropriate restriction enzymes to yield 11 fragments, suitable for use as molecular weight standards for agarose gel electrophoresis. The DNA includes fragments ranging from 100-1,500 base pairs. The 500 and 1,500 base pair bands have increased intensity to serve as reference points.\u00a0The approximate mass of DNA in each band is provided (0.5 \u03bcg a load) for approximating the mass of DNA in comparably intense samples of similar size.<\/p>\n

Source\u00a0:\u00a0<\/strong>PCR products and double-stranded DNA digested with appropriate restriction enzymes are phenol extracted and equilibrated to 10 mM Tris-HCl (pH 8.0) and 10mM EDTA.<\/p>\n

Reaction Setup<\/strong>:<\/strong>\u00a0<\/strong>5 \u03bcl \/ well (\u00a0** Containing orange G & xylene cyanol FF as tracking dyes. )<\/p>\n

Range:\u00a0<\/strong>100 – 1,500 bp<\/p>\n

Number of\u00a0<\/strong>bands:<\/strong>\u00a0<\/strong>11
\nStorage :<\/strong><\/p>\n

Stable for up to 6 months at 25\u00b0C.
\nStable for up to 12 months at 4\u00b0C.
\nStable for up to 24\u00a0months at -20\u00b0C.<\/p>\n

\u5efa\u8b70\u4f7f\u7528\u91cf<\/span><\/strong>\u00a05 \u03bcl \/ well (\u00a0** Containing orange G & xylene cyanol FF as tracking dyes. )<\/span>
\nSource\u00a0:\u00a0PCR products and double-stranded DNA digested with appropriate restriction enzymes are phenol extracted and equilibrated to 10 mM Tris-HCl (pH 8.0) and 10mM EDTA.<\/span><\/p>\n

\"\u7db2\u7ad9\u898f\u683c\u5c0f\u5716<\/p>\n

\u85cd\u5149\u89c0\u5bdf\u7bb1(\u542b\u767d\u5149)<\/strong><\/a>\u00a0|\u00a0Mupid-2\u00a0<\/strong>\u6c34\u5e73\u96fb\u6cf3\u69fd<\/strong><\/a>\u00a0|<\/p>\n

\"BIO-HELIX<\/a>\"MUPID<\/a><\/p>\n

\"HOT\"<\/a>BluPAD \u96d9 LED \u85cd\/\u767d\u5149\u900f\u5c04\u5100 | BIO-HELIX \u8ca8\u865fBP001CU\u00a0\u85cd\u5149\u89c0\u5bdf\u7bb1(\u542b\u767d\u5149)<\/strong><\/span><\/p>\n

\"\u85cd\u5149LED\u900f\u7167\u5668\uff5c\u767d\u5149LED\u900f\u7167\u5668\"<\/p>\n

\"\"<\/p>\n

\u2022\u85cd\u5149\u6a21\u5f0f<\/strong><\/span><\/p>\n

\u4ee5470nm LED\u71c8\u70ba\u6fc0\u767c\u5149\u6e90\uff0c\u63a1\u7528\u85cd\u5149\u6a21\u5f0f\u89c0\u5bdf\u4f7f\u7528\u7192\u5149\u67d3\u8272\u8a66\u5291\u7684\u5b9a\u6027\u548c\u5b9a\u91cf\u6838\u9178\u6216\u86cb\u767d\u8cea\u5be6\u9a57\u3002\u9664\u4e86\u517c\u5bb9\u6211\u5011\u5b89\u5168\u7684\u8a66\u5291\uff0c\u5982Novel Juice\u3001Novel Green\u3001Novel Green Plus\u3001OnePCR\u7cfb\u5217\u3001Nimble Juice\u548cNimble Juice R-TYPE\uff0c\u5b83\u9084\u8207\u5e02\u5834\u4e0a\u7684\u5927\u591a\u6578\u7192\u5149\u67d3\u8272\u7522\u54c1\u517c\u5bb9\uff0c\u4f8b\u5982 SYBR Gold\u3001SYBR Green I & II\u3001SYPRO Ruby\u3001SYPRO Orange \u548c Coomassie Fluor Orange \u67d3\u8272\u5291\u3001GelStar\u3001GelGreen\u3002\u4e09\u500b\u4e0d\u540c\u7d1a\u5225\u7684\u5149\u5f37\u8abf\u7bc0\u529f\u80fd\u4f7f\u7528\u6236\u53ef\u4ee5\u6839\u64da\u6a23\u54c1\u6fc3\u5ea6\u9032\u884c\u5149\u5f37\u5c0d\u6bd4\u5ea6\u8abf\u7bc0\uff0c\u4ee5\u5be6\u73fe\u6700\u4f73\u6210\u50cf\u8cea\u91cf\u3002\u6b64\u5916\uff0c\u78c1\u6027\u7425\u73c0\u8272\u904e\u6ffe\u5668\uff0c<\/p>\n

\"\"<\/p>\n

\u25bc\u00a0\u5de6\uff1a\u51dd\u81a0\u5207\u5272 \/ \u53f3\uff1a\u6210\u50cf<\/span><\/strong><\/span><\/p>\n

\"\u8d85\u8584\u85cd\/\u767d<\/p>\n

\u2022\u767d\u5149\u6a21\u5f0f<\/strong><\/span><\/p>\n

White Light\u6a21\u5f0f\u4f7f\u7528\u5168\u6ce2\u9577\u767d\u5149LED\u4f5c\u70ba\u6fc0\u767c\u5149\u6e90\uff0c\u8868\u73fe\u51fa\u67d4\u548c\u5747\u52fb\u7684\u7279\u9ede\uff0c\u9069\u7528\u65bc\u5c0d\u8003\u99ac\u65af\u85cd\u6216\u9280\u67d3\u7684SDS-PAGE\u51dd\u81a0\u9032\u884c\u89c0\u5bdf\u6216\u6210\u50cf\u3002\u5b83\u4e5f\u53ef\u4ee5\u7528\u4f5c\u7c21\u55ae\u7684\u96fb\u5f71\u89c0\u5bdf\u900f\u7167\u5668\uff0c\u7528\u65bc\u6aa2\u67e5\u7528\u65bc\u7814\u7a76\u6216\u81e8\u5e8a\u76ee\u7684\u7684 X \u5c04\u7dda\u81a0\u7247\u3002\u5177\u6709\u5149\u5f37\u8abf\u7bc0\u529f\u80fd\uff0c\u53ef\u6839\u64da\u89c0\u5bdf\u8981\u6c42\u5c0d\u5149\u5f37\u9032\u884c\u4e09\u7d1a\u5c0d\u6bd4\u5ea6\u8abf\u7bc0\uff0c\u4ee5\u9054\u5230\u6700\u4f73\u6210\u50cf\u8cea\u91cf\u3002<\/p>\n

\"\u767d\u5149LED\u900f\u7167\u5100\uff5cSDS-PAGE,<\/p>\n

\u2022\u5be6\u5730\u5b78\u7fd2\u6a21\u5f0f<\/strong><\/p>\n

\u53ef\u8207\u5916\u90e8\u79fb\u52d5\u96fb\u6e90\u9023\u63a5\uff0c\u9032\u884c\u7121\u6182\u7684\u73fe\u5834\u5be6\u9a57\u3002
\n*\u79fb\u52d5\u96fb\u6e90\u4e0d\u5305\u542b\u5728\u5305\u88dd\u5167\uff0c\u9700\u8981\u7531\u7528\u6236\u8cfc\u8cb7\u3002<\/p>\n

\u2022 \u96d9\u5149\u6e90<\/strong>
\nLED\u767d\u5149\u548c\u85cd\u5149\uff1a\u5728\u57fa\u790e\u79d1\u5b78\u548c\u91ab\u5b78\u8a3a\u65b7\u7814\u7a76\u9818\u57df\u5177\u6709\u5ee3\u6cdb\u7684\u9069\u7528\u6027\u548c\u517c\u5bb9\u6027\u3002<\/p>\n

\u2022 \u78c1\u6027\u904e\u6ffe\u5668<\/strong>
\n\u7121\u9278\u93c8\u8a2d\u8a08\u70ba\u7528\u6236\u63d0\u4f9b\u7c21\u55ae\u3001\u5b89\u5168\u548c\u65b9\u4fbf\uff0c\u4e0d\u6703\u5c0d\u9278\u93c8\u9644\u8fd1\u7684\u904e\u6ffe\u5668\u9020\u6210\u4efb\u4f55\u640d\u58de\u3002\u9032\u884c\u89c0\u5bdf\u548c\u51dd\u81a0\u5207\u5272\u4e0d\u9700\u8981\u8b77\u76ee\u93e1\u3002<\/p>\n

\u2022 Bottom-Up LED \u7167\u660e<\/strong>
\n\u907f\u514d\u4e86\u5074\u9762\u7167\u660e\u5f15\u8d77\u7684\u53cd\u5c04\u5149\u7684\u5e72\u64fe\uff0c\u63d0\u9ad8\u4e86\u89c0\u5bdf\u548c\u6210\u50cf\u8cea\u91cf\u3002\u7531\u65bc LED \u71c8\u7d93\u4e45\u8010\u7528\u4e14\u5b89\u5168\uff0c\u4e0d\u6703\u50cf\u901a\u5e38\u5728\u7d2b\u5916\u7dda\u7167\u5c04\u4e0b\u90a3\u6a23\u5c0d\u773c\u775b\u548c\u76ae\u819a\u6216\u5be6\u9a57\u6a23\u54c1\u9020\u6210\u50b7\u5bb3\u3002<\/p>\n

\u2022 \u5149\u5f37\u53ef\u8abf\uff083\u7d1a\uff09<\/strong>
\n\u6839\u64da\u6a23\u54c1\u6578\u91cf\u6216\u89c0\u5bdf\u8981\u6c42\u8abf\u7bc0\u5149\u5f37\u548c\u5c0d\u6bd4\u5ea6\uff0c\u53ef\u9054\u5230\u6700\u4f73\u7684\u89c0\u5bdf\u6216\u6210\u50cf\u8cea\u91cf\u3002<\/p>\n

\u2022 5 \u5206\u9418\u81ea\u52d5\u95dc\u6a5f<\/strong>
\n\u4fdd\u8b77\u900f\u7167\u5100\u514d\u53d7\u56e0\u7528\u6236\u64cd\u4f5c\u758f\u5ffd\u9020\u6210\u7684\u98a8\u96aa\u3002<\/p>\n

\u2022 \u8a2d\u8a08\u5e2b\u8a2d\u8a08\u7684\u91d1\u5c6c\u5916\u6bbc<\/strong>
\n\u900f\u7167\u5100\u7684\u5e95\u5ea7\u66f4\u52a0\u7a69\u5b9a\uff0c\u5f9e\u800c\u7c21\u5316\u4e86\u64cd\u4f5c\u904e\u7a0b\u3002<\/p>\n

\u2022 \u7cbe\u5de7\u7dca\u6e4a\u7684\u8a2d\u8a08<\/strong>
\n\u4fbf\u65bc\u79fb\u52d5\u548c\u5b58\u653e\uff0c\u9069\u5408\u91ce\u5916\u8003\u5bdf\u6642\u7684\u5be6\u9a57\u89c0\u5bdf\u3002<\/p>\n

\u2022 \u4f7f\u7528\u79fb\u52d5\u96fb\u6e90\u589e\u5f37\u4fbf\u651c\u6027<\/strong>
\n\u8f15\u9b06\u9032\u884c\u6236\u5916\u5be6\u9a57\u3002
\n**\u79fb\u52d5\u96fb\u6e90\u4e0d\u5305\u542b\u5728\u5305\u88dd\u5167\uff0c\u9700\u8981\u7531\u7528\u6236\u8cfc\u8cb7\u3002<\/p>\n

BluPAD \u96d9 LED \u85cd\u5149\/\u767d\u5149\u900f\u5c04\u5100<\/strong><\/span><\/p>\n

BluPAD\u662f\u4e00\u6b3e\u5275\u65b0\u8a2d\u8a08\u7684\u96d9\u5149\u6e90\u900f\u7167\u5100\uff0c\u9069\u7528\u65bc\u751f\u547d\u79d1\u5b78\u7814\u7a76\u7684\u5404\u500b\u9818\u57df\uff0c\u5c0d\u6838\u9178\u548c\u86cb\u767d\u8cea\u9032\u884c\u89c0\u5bdf\u548c\u5206\u6790\u3002\u5176\u91dd\u5c0d\u51dd\u81a0\u5207\u5272\u3001\u6578\u64da\u6210\u50cf\u548c\u6b78\u6a94\u7b49\u5f8c\u671f\u89c0\u5bdf\u61c9\u7528\u7684\u6700\u512a\u5316\u548c\u4eba\u6027\u5316\u8a2d\u8a08\uff0c\u5e0c\u671b\u70ba\u7814\u7a76\u4eba\u54e1\u63d0\u4f9b\u5168\u65b0\u7684\u8212\u9069\u3001\u65b9\u4fbf\u548c\u5b89\u5168\u7684\u9ad4\u9a57\u3002<\/p>\n