生物技术
生物技術
생물기술
BIOTECHNOLOGY
2009年
6期
57-61
,共5页
DNA%毛细管电泳%紫外检测%水柱法场扩大堆积注射%压力%灵敏度热
DNA%毛細管電泳%紫外檢測%水柱法場擴大堆積註射%壓力%靈敏度熱
DNA%모세관전영%자외검측%수주법장확대퇴적주사%압력%령민도열
DNA%capillary electrophoresis%ultraviolet detection%water-plug field-amplified stacking injection pressure%sensitivity
目的:采用水柱法场扩大堆积技术,提高毛细管电泳紫外分析核酸灵敏度.方法:已知浓度的DNA Marker为标准样品,TE缓冲液递度稀释,压力进样前加一段去离子水柱(0.5psi,20s),观察不同浓度下压力进样紫外检测图谱.结果:在对分离度无明显影响下,将压力进样时间延长至0.5psi,990s,与常规的压力进样(0.5psi,10s)相比,灵敏度提高了94.4倍.与时间延长后的压力进样(0.5psi,90s)相比,灵敏度提高8.2倍,最低检测总浓度为1ng/μl,DNA的检测限降至80ng/ml(S/N=3),比前人报道的7ng/μl检测限提高了87.5倍.结论:验证了水柱法场扩大堆积注射可以有效提高毛细管电泳紫外检测核酸灵敏度.
目的:採用水柱法場擴大堆積技術,提高毛細管電泳紫外分析覈痠靈敏度.方法:已知濃度的DNA Marker為標準樣品,TE緩遲液遞度稀釋,壓力進樣前加一段去離子水柱(0.5psi,20s),觀察不同濃度下壓力進樣紫外檢測圖譜.結果:在對分離度無明顯影響下,將壓力進樣時間延長至0.5psi,990s,與常規的壓力進樣(0.5psi,10s)相比,靈敏度提高瞭94.4倍.與時間延長後的壓力進樣(0.5psi,90s)相比,靈敏度提高8.2倍,最低檢測總濃度為1ng/μl,DNA的檢測限降至80ng/ml(S/N=3),比前人報道的7ng/μl檢測限提高瞭87.5倍.結論:驗證瞭水柱法場擴大堆積註射可以有效提高毛細管電泳紫外檢測覈痠靈敏度.
목적:채용수주법장확대퇴적기술,제고모세관전영자외분석핵산령민도.방법:이지농도적DNA Marker위표준양품,TE완충액체도희석,압력진양전가일단거리자수주(0.5psi,20s),관찰불동농도하압력진양자외검측도보.결과:재대분리도무명현영향하,장압력진양시간연장지0.5psi,990s,여상규적압력진양(0.5psi,10s)상비,령민도제고료94.4배.여시간연장후적압력진양(0.5psi,90s)상비,령민도제고8.2배,최저검측총농도위1ng/μl,DNA적검측한강지80ng/ml(S/N=3),비전인보도적7ng/μl검측한제고료87.5배.결론:험증료수주법장확대퇴적주사가이유효제고모세관전영자외검측핵산령민도.
Objective: Enhancing the UV detection sensitivity of capillary electrophoresis for DNA by water-plug field-amplified stacking injection . Method: The known concentration of DNA marker were as standard samples, TE buffer diluted the standard samples gardiently, inserted a water-plug(0.5psi, 20s)before pressure injection, observed the CE-UV map in different concentration.Result: The pressure injection time was prolonged until 0.5psi, 990s without deteriorating the resolution, compared to normal pressure injection(0.5psi, 10s)and extended pressure injec-tion(0.5psi, 90s), the sensitivity was enhanced 94.4 folds and 8.2 folds respectively. 1ng/μl was the lowest detection overall concentration of DNA, 80ng/ml(S/N = 3)is the detection limit, the result was 87.5 folds better than the former researchers' detection limit of 7 ng/μl. Conclusion: The experiment validated the water-plug field-amplified stacking injection was an effective approach to enhance the CE-UV detection sensitivity of DNA.
