Southern Blot
Southern印锟紼锟絪锟斤拷锟斤拷Southern blot锟斤拷锟斤拷1975锟斤拷锟斤拷英锟斤拷(gu锟斤拷)锟斤拷southern锟斤拷(chu锟斤拷ng)锟斤拷,锟斤拷锟斤拷锟叫撅拷DNA锟紻锟絍锟侥伙拷锟斤拷锟斤拷锟絞(sh锟斤拷)锟斤拷Southern印锟紼锟絪锟斤拷锟斤拷Southern blot锟斤拷锟斤拷锟組(j锟斤拷n)锟叫伙拷锟斤拷MDNA锟截讹拷锟斤拷锟叫讹拷位锟斤拷通锟矫凤拷锟斤拷,锟斤拷一锟斤拷锟斤拷锟矫拷脂锟斤拷锟斤拷锟絲锟接撅拷锟斤拷x锟斤拷(j锟斤拷ng)锟斤拷锟斤拷锟皆冿拷(n锟斤拷i)锟斤拷酶锟斤拷锟斤拷锟斤拷DNA片锟斤拷,锟斤拷锟斤拷锟絲锟较碉拷DNA變锟皆诧拷锟斤拷原位锟斤拷锟斤拷锟紻NA片锟斤拷锟紻(zhu锟斤拷n)锟斤拷锟斤拷锟斤拷锟斤拷膜锟斤拷锟斤拷锟斤拷锟斤拷锟斤拷支锟斤拷锟斤拷锟较o拷锟斤拷(j锟斤拷ng)锟缴匡拷锟斤拷锟斤拷锟斤拷锟解線锟斤拷锟斤拷潭锟,锟斤拷锟斤拷锟絚锟斤拷锟斤拷(du锟斤拷)锟斤拷(y锟斤拷ng)锟結(ji锟斤拷)锟斤拷(g锟斤拷u)锟侥橈拷(bi锟斤拷o)記探锟斤拷M(j锟斤拷n)锟斤拷锟絪锟斤拷,锟斤拷锟矫凤拷锟斤拷锟斤拷锟紷影锟斤拷酶锟斤拷锟斤拷(y锟斤拷ng)锟紷色锟斤拷锟侥讹拷锟絲锟統(c锟斤拷)锟截讹拷DNA锟斤拷锟接的猴拷锟斤拷,锟斤拷锟斤拷锟絑锟斤拷锟絖(d锟斤拷)锟结供DIG锟斤拷(bi锟斤拷o)記锟斤拷Southern Blot锟斤拷锟絞(sh锟斤拷)锟斤拷锟斤拷(w锟斤拷),锟斤拷
锟斤拷锟斤拷(w锟斤拷)要锟斤拷
1锟斤拷锟斤拷锟絪锟斤拷目锟斤拷(bi锟斤拷o)锟斤拷锟斤拷锟斤拷锟叫和猴拷锟叫达拷锟絪锟斤拷锟斤拷锟斤拷锟斤拷(y锟斤拷ng)锟斤拷锟絴(zh锟斤拷)锟斤拷,锟斤拷锟揭傦拷锟斤拷(hu锟斤拷)锟斤拷锟斤拷(j锟斤拷)Southern blot锟斤拷(sh锟斤拷)锟(y锟斤拷n)要锟斤拷锟斤拷锟斤拷锟絰锟斤拷全锟絃(zh锟斤拷ng)锟津部分區(q锟斤拷)锟斤拷锟斤拷锟斤拷探锟斤拷锟斤拷锟,锟斤拷使锟斤拷PCR锟斤拷锟斤拷(bi锟斤拷o)記锟截革拷锟斤拷探針★拷
2,锟斤拷锟斤拷锟絪锟斤拷锟侥革拷锟絴(zh锟斤拷)锟斤拷DNA锟斤拷品锟絴(zh锟斤拷)锟斤拷锟斤拷锟絇(gu锟斤拷n)锟斤拷要,锟斤拷锟絈锟斤拷锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟侥成旓拷锟斤拷請(q锟斤拷ng)锟絖锟斤拷DNA锟斤拷锟斤拷锟絴(zh锟斤拷)锟斤拷,锟斤拷锟組锟斤拷锟斤拷锟斤拷锟組(j锟斤拷n)锟斤拷試锟斤拷锟斤拷锟斤拷取锟斤拷委锟斤拷锟斤拷锟絑锟斤拷锟絖(d锟斤拷)锟斤拷取,锟斤拷
A锟斤拷锟斤拷锟斤拷MDNA锟絲锟統(c锟斤拷)锟斤拷OD260/OD280=1.8-2.0,锟斤拷锟斤拷却锟斤拷锟0.3锟斤拷g/ul,锟斤拷锟絪锟斤拷锟斤拷模锟斤拷DNA锟絪60 锟斤拷g/泳锟斤拷/锟轿o拷
B,锟斤拷請(q锟斤拷ng)?zh锟斤拷)峁╋拷锟斤拷锟組DNA锟絲锟統(c锟斤拷)锟紻片,锟斤拷要锟斤拷o(w锟斤拷)锟斤拷锟解,锟給(w锟斤拷)锟斤拷锟斤拷锟斤拷染,锟斤拷
C锟斤拷锟絲锟統(c锟斤拷)锟斤拷品锟酵滐拷锟絓(y锟斤拷n)輸锟斤拷锟斤拷司,锟斤拷
3,锟斤拷锟酵戯拷锟斤拷锟斤拷o(w锟斤拷)锟斤拷锟结供锟斤拷品锟絴(zh锟斤拷)锟侥伙拷锟斤拷MDNA锟斤拷品锟斤拷锟斤拷锟絑锟斤拷锟絖(d锟斤拷)锟斤拷锟皆庯拷锟斤拷锟狡傦拷锟斤拷锟斤拷锟絊outhern Blot锟絲锟統(c锟斤拷)锟斤拷DNA锟斤拷品,锟斤拷锟酵戯拷锟结供锟斤拷品要锟斤拷锟斤拷锟铰o拷
A,锟斤拷锟斤拷液锟斤拷锟斤拷锟斤拷锟斤拷500ul锟斤拷锟斤拷锟斤拷锟斤拷锟竭干憋拷锟絓(y锟斤拷n)輸
B,锟斤拷锟組锟斤拷锟斤拷锟斤拷锟斤拷锟斤拷小锟斤拷2g,锟缴憋拷锟絓(y锟斤拷n)輸
C,锟斤拷锟斤拷(x锟斤拷)锟斤拷锟斤拷锟结供锟斤拷锟絩锟侥硷拷(x锟斤拷)锟斤拷锟斤拷锟斤拷,锟斤拷锟組锟斤拷锟斤拷要使锟斤拷锟絵锟斤拷液锟絖洗
4锟斤拷請(q锟斤拷ng)注锟斤拷锟斤拷锟斤拷MDNA酶锟叫凤拷锟斤拷,锟斤拷锟斤拷锟斤拷(xi锟斤拷ng)目锟斤拷锟斤拷要锟矫碉拷锟角筹拷規(gu锟斤拷)锟斤拷锟斤拷锟斤拷锟皆冿拷(n锟斤拷i)锟斤拷酶,锟斤拷锟斤拷锟絶锟斤拷锟斤拷锟矫革拷锟斤拷M(f锟斤拷i)锟矫伙拷锟叫客戯拷锟结供锟斤拷锟斤拷(y锟斤拷ng)锟斤拷锟斤拷锟斤拷锟皆冿拷(n锟斤拷i)锟斤拷酶锟斤拷
锟斤拷锟斤拷(w锟斤拷)锟斤拷锟斤拷锟斤拷锟斤拷
1.锟斤拷锟斤拷MDNA锟斤拷锟,锟斤拷锟斤拷锟斤拷锟斤拷要锟斤拷,锟斤拷
2.探锟斤拷O(sh锟斤拷)計(j锟斤拷)锟较成o拷锟斤拷锟斤拷锟揭拷锟斤拷锟
3. DIG探針橈拷(bi锟斤拷o)記,锟斤拷
4. Southern锟絪锟斤拷锟絲锟統(c锟斤拷),锟斤拷
5. 锟结交锟斤拷影锟絲锟統(c锟斤拷)锟斤拷(b锟斤拷o)锟芥,锟斤拷锟斤拷詳锟斤拷(x锟斤拷)锟斤拷锟斤拷锟斤拷锟斤拷蛯锟(sh锟斤拷)锟(y锟斤拷n)锟結(ji锟斤拷)锟斤拷锟斤拷,锟斤拷
Southern Blot锟斤拷锟斤拷(w锟斤拷)锟斤拷锟斤拷锟侥硷拷
锟斤拷锟絑锟斤拷锟絖(d锟斤拷)負(f锟斤拷)責(z锟斤拷)锟斤拷(du锟斤拷)锟酵戯拷锟结供锟斤拷品锟組(j锟斤拷n)锟斤拷DIG探針橈拷(bi锟斤拷o)記锟絚Southern锟絪锟斤拷锟絲锟統(c锟斤拷)锟斤拷锟絞(sh锟斤拷)锟斤拷锟斤拷(w锟斤拷),锟斤拷锟斤拷K锟结供锟絲锟統(c锟斤拷)锟斤拷(b锟斤拷o)锟芥及锟絲锟統(c锟斤拷)锟紻片
1锟斤拷DNA锟斤拷品锟絲锟統(c锟斤拷)锟斤拷(b锟斤拷o)锟芥(锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷品锟斤拷锟斤拷還,锟斤拷請(q锟斤拷ng)注锟解保锟斤拷锟捷o拷
2,锟斤拷锟斤拷(bi锟斤拷o)锟斤拷(zh锟斤拷n)锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷锟教堬拷(b锟斤拷o)锟斤拷
3锟斤拷southern blot锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟結(ji锟斤拷)锟斤拷锟絲锟統(c锟斤拷)锟斤拷(b锟斤拷o)锟斤拷
4,锟斤拷锟斤拷锟斤拷菍锟(sh锟斤拷)锟(y锟斤拷n)锟斤拷锟絶锟斤拷锟揭拷锟,锟斤拷請(q锟斤拷ng)?zh锟斤拷)锟角帮拷锟诫娀锟斤拷]锟斤拷锟絝(shu锟斤拷)锟斤拷
锟斤拷(sh锟斤拷)锟(y锟斤拷n)锟斤拷锟斤拷锟紻片锟斤拷

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