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Elkin 绛変娇鐢ㄧ晶鍖栫鐝犲垎绂荤函鍖栬川绮扗NA銆傝娉曞湪缁嗚優瑁傝В鍚庯紝绂诲績鍒嗙鍚川绮掔殑姘寸浉锛屽啀鍔犲叆缇у寲鐨勭绮掞紝鐒跺悗鐢 PEG/ NaCl 娌夋穩锛屼娇鐩殑DNA 鍚搁檮鑷崇鐝狅紝鏈€鍚庣鍦哄垎绂昏鍚搁檮鐨凞NA 锛岀粡涔欓唶娲楁钉锛岀敤姘存礂鑴憋紝鍙幏寰楅珮浜ч噺鐨勯€傜敤浜庢瘺缁嗙娴嬪簭鐨勬ā鏉緿NA銆侟/span>

涔熸湁鐢ㄩ搧绮掍负鍥虹浉鏀寔鐗╋紝缁忕鍦哄垎绂昏€岀函鍖栬川绮扗NA 鐨勬姤閬撱€傜粏鑿岀敤婧惰弻閰?鐓哺娉曡瑙ｏ紝璐ㄧ矑琚噴鏀捐嚦鎮诞娑蹭腑锛 鍔犻搧鐝犳崟鑾凤紝鐢ㄧ鍦轰娇閾佺彔鍒嗙锛岀粡婕傛礂鍚庣敤姘存礂鑴辫川绮掞紝鍙幏寰楅珮浜ч噺銆佹祴搴忕骇鐨勮川绮扗NA銆侟/span>

浜插拰灞傛瀽鏄埄鐢ㄥ緟鍒嗙鐗╄川涓庡畠浠殑鐗瑰紓鎬ч厤浣撻棿鎵€鍏锋湁鐨勭壒寮傛€т翰鍜屽姏鏉ュ垎绂荤墿璐ㄧ殑涓€绫诲眰鏋愭柟娉曘€侰handler 绛夋姤閬撲簡涓€绉嶇敤鑲芥牳閰?PNA)鍒嗙鏍搁吀鐨勬柟娉曘€侾NA 鏄竴绫讳互 N-(2-姘ㄤ箼鍩?2鐢樻皑閰哥粨鏋勫崟鍏冧负楠ㄦ灦鐨凞NA 绫讳技鐗╋紝鍙綔涓虹函鍖栫毊鍏?pg)绾ф牳绯栦綋DNA(rDNA)鍜屾牳绯栦綋RNA(rRNA)鐨勮瘯鍓傘€傚湪璇ユ柟娉曚腑锛屼互鐢熺墿绱犳爣璁扮殑鑲芥牳閰?peptide nucleic acids 锛孭NAs)涓烘帰閽堬紝浠ュ寘琚簡鎶楃敓铔嬬櫧閾捐弻绱犵殑纾佺彔浣滀负鍥虹浉杞戒綋銆侾NA 鎺㈤拡鍦ㄩ珮鐩愮幆澧冧笅锛 涓庣洰鐨勬牳閰?DNA 鎴 RNA)娣峰悎锛岀粡鐓哺銆佸啺娴淬€佹俯鑲叉潅浜ゆ楠ゅ悗锛岀洿鎺ュ姞鍏ュ寘琚簡鎶楃敓铔嬬櫧閾捐弻绱犵殑椤虹鎬ч绮掞紝缁忛潤缃崟鑾稰NA-鏍搁吀鏉備氦浣擄紝姘存礂鑰岃幏寰楃函鍖栫殑鏍搁吀銆侟/span>

Schluep 绛変害鍩轰簬浜插拰灞傛瀽鍘熺悊锛岄噰鐢ㄤ竴绉嶄笁铻烘棆浣揇NA 鐨勬柟寮忚繘琛岃川绮扗NA 鐨勫垎绂汇€備笁铻烘棆浣揇NA 鐢卞悓璐ㄥ槍鍛ゃ€佸悓璐ㄥ槯鍟跺弻铻烘棆閾句笌鍚岃川鍢у暥鍗曢摼缁勬垚锛屽崟閾句笂鐨凾 璇嗗埆A 路T 纰卞熀锛屽褰㈡垚T 路A 路T 涓夎仈浣擄紝璐ㄥ瓙鍖栫殑鍗曢摼鑳炲槯鍟?C+)璇嗗埆G路C 纰卞熀瀵瑰舰鎴怌 路G路C 涓夎仈浣撱€傚湪閫傚綋鐨勬潯浠朵笅锛屼笁铻烘棆浣撶殑缁撳悎鍏锋湁楂樼壒寮傛€у拰楂樼ǔ瀹氭€с€傚皢閰嶄綋鑱氬槯鍟跺鏍歌嫹閰搁摼閫氳繃鍖栧鏂规硶杩炴帴鑷砈ephacryl S21000 SF 棰楃矑涓婂舰鎴愪翰鍜岃浇浣撱€傚綋鍚洰鐨勫簭鍒楃殑璐ㄧ矑DNA 婧舵恫涓庡叾娣峰悎鏃讹紝鍦ㄩ吀鎬х幆澧?p H 4.5锝?.5)涓嬶紝璐ㄧ矑缁撳悎鑷充翰鍜岃浇浣撻绮掍笂锛屾憾娑蹭腑楂樻祿搴︾殑NaCl 鍙ǔ瀹氫笁鑱斾綋褰㈠紡骞跺噺灏戜笌铔嬬櫧璐ㄣ€佺粏鑳濪NA 鐨勯潪鐗瑰紓鎬х粨鍚堛€傜粡涓€娈垫椂闂村弽搴斿悗锛岄绮掓偓娴恫琚姞鑷充竴灞傛瀽鏌憋紝鐢ㄩ€傚綋鐨勬礂鑴辨恫鏀瑰彉p H 鍊艰嚦纰辨€х幆澧冿紝鍙娇涓夎仈浣撹В鑱氾紝璐ㄧ矑琚礂鑴便€傜粡璇ユ硶鍒嗙璐ㄧ矑DNA锛岃川绮掍骇閲忓彲杈惧埌鍔犲叆閲忕殑62%銆侟/span>

涔熸湁鐢⊿chizophyllan(SPG)鍒跺浜插拰灞傛瀽鏌卞垎绂荤函鍖 RNA 鐨勬姤閬撱€係PG鏄竴绉嵨?1 锛?2钁¤仛绯栵紝鍦ㄤ綆娓╀笅锛屽惈RNA 鐨勬祦鍔ㄧ浉閫氳繃灞傛瀽鏌憋紝Poly(C)鍜宲oly(A)涓嶴PG閫氳繃姘㈤敭鍜岀枏姘翠綔鐢ㄥ舰鎴愬鍚堢墿鑰岃鍚搁檮浜庢煴涓婏紝鐒跺悗閫氳繃鏀瑰彉缂撳啿娑叉垚鍒嗭紝灏嗚鍚搁檮鐨凴NA 娲楄劚銆備翰鍜屽眰鏋愬簲鐢ㄤ簬鏍搁吀鍒嗙涓庣函鍖栫殑鍙︿竴涓緥瀛愭槸鐢╫ligo(dT)2绾ょ淮绱犲眰鏋愭硶浠庣湡鏍哥粏鑳炴€ RNA 涓垎绂诲甫poly(A)灏剧殑mRNA銆傚湪璇ユ柟娉曚腑锛岀煭閾緊ligo(dT)閫氳繃鍏?2纾烽吀涓庣氦缁寸礌鐨勭緹鍩哄叡浠风粨鍚堣€岃繛鎺ヨ嚦绾ょ淮绱犱粙璐ㄤ笂銆傚綋鏍锋湰缁忚繃oligo(dT)鏌辨椂锛宮RNA 鍥犲叾poly(A)鍙笌鐭摼oligo(dT)褰㈡垚绋冲畾鐨凴NA2DNA 鏉傚悎閾撅紝鑰岃杩炴帴鍒扮氦缁寸礌浠嬭川涓婏紝浠庤€屼笌鍏朵粬RNA 鍒嗙銆傚湪閫傚綋鐨勬潯浠朵笅(浣庣洂銆佸姞鐑?锛宲oly(A)RNA 鍙姘存礂鑴辫€屽緱浠ョ函鍖栥€侟/span>

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(3)瀵嗗害姊害绂诲績娉旤br/> 瀵嗗害姊害绂诲績涔熺敤浜庢牳閰哥殑鍒嗙鍜屽垎鏋愩€傚弻閾綝NA銆佸崟閾綝NA銆丷NA 鍜岃泲鐧借川鍏锋湁涓嶅悓鐨勫瘑搴︼紝鍥犺€屽彲缁忓瘑搴︽搴︾蹇冨舰寮忓舰鎴愪笉鍚屽瘑搴︾殑绾牱鍝佸尯甯︼紝 璇ユ硶閫傜敤浜庡ぇ閲忔牳閰告牱鏈殑鍒跺锛屽叾涓隘鍖栭摨銆佹捍鍖栦箼閿搴﹀钩琛＄蹇冩硶琚涓烘槸绾寲澶ч噺璐ㄧ矑DNA 鐨勯閫夋柟娉曘€傛隘鍖栭摨鏄牳閰稿瘑搴︽搴︾蹇冪殑鏍囧噯浠嬭川锛屾搴︽恫涓殑婧村寲涔欓敪涓庢牳閰哥粨鍚堬紝绂诲績鍚庡舰鎴愮殑鏍搁吀鍖哄甫缁忕传澶栫伅鐓у皠锛屼骇鐢熻崸鍏夎€岃妫€娴嬶紝鐢ㄦ敞灏勯拡澶寸┛鍒哄洖鏀跺悗锛岄€氳繃閫忔瀽鎴栦箼閱囨矇娣€闄ゅ幓姘寲閾€岃幏寰楃函鍖栫殑鏍搁吀銆侟/span>