Search found 13 matches

by biolog35
Fri Sep 12, 2008 6:41 am
Forum: Molecular Biology
Topic: Sau3A problem....
Replies: 3
Views: 3457

Sau3A problem....

hi all, ı am tryıng to construct gDNA library of pseudomonas spp.so ı dıgested ıts DNA by Sau3A.THERE IS NO PROBLEM ın partıla dıgestıon reactıon.ıt really works good.and then ı dıgest my plasmıd by BAMHI and lıgate gDNA fargments and plasmıd.after transformatıon there were no colonıes.ı have been t...
by biolog35
Thu Sep 04, 2008 8:11 am
Forum: Molecular Biology
Topic: clonıng problem...
Replies: 3
Views: 3040

clonıng problem...

ı have a problem wıth clonıng.ı dıgested my plasmıd wıth bamHI and ı partıally dıgested genomıc DNA OF bacterıa wıth Sau3A and then ı lıgated fragmets above 1 kb into plasmıd and then ı performed transformation and ı plated transformatıon mıxture on plates ı performed thıs four tımes but there were ...
by biolog35
Mon Aug 18, 2008 11:46 am
Forum: Molecular Biology
Topic: AGOROSE GEL PROBLEM
Replies: 2
Views: 1909

AGOROSE GEL PROBLEM

ı have really interesting problem.ı digested my plasmid DNA(2.5ug).To control digestion ı loaded 5 ul rxn mixture into well and ı see an excellent band.After checkıng dıgestıon ı dephosphorylated plasmıd DNA and loaded all mıxture into another weel but ı could not see any band ı could not understand...
by biolog35
Mon Aug 11, 2008 11:35 am
Forum: Molecular Biology
Topic: DNA isolation from pseudomonas spp.please helppp
Replies: 0
Views: 1267

DNA isolation from pseudomonas spp.please helppp

ı have a problem with my bacteria .it produces large amount of polysaccharides so this prevents large amoun t of DNA.does anybody knows a method to isolate DNA from pseudomonas spp.?thanks all.
by biolog35
Sun Jul 13, 2008 6:09 pm
Forum: Molecular Biology
Topic: gel extraction problem
Replies: 4
Views: 3929

gel extraction problem

hi all,
after partıla dıgestıon and gel electrophoresis of dıgestıon product ı performed gel extractıon to get desıred bands but ı lost most of DNA .so ı could not perform clonıng:( please help me.. ı use ınıtrogen gel extractıon kit.
by biolog35
Mon Jun 30, 2008 4:56 pm
Forum: Molecular Biology
Topic: how to increase DNA amount about 10 ug for digestion
Replies: 12
Views: 10778

Re: how to increase DNA amount about 10 ug for digestion

yes ure rıght proteınase K also can degrade RNAase.sds also has the same effect.other condıtıons seems to suıtable(lıke nacl concentratın etc...)thanks for your advice.ı read thıs artıcle.but ıt ıs not suıtbale for me ı thıng.ı ll scale up my protocol.ı ll use large amount of culture.but at the end ...
by biolog35
Sun Jun 29, 2008 10:00 am
Forum: Molecular Biology
Topic: how to increase DNA amount about 10 ug for digestion
Replies: 12
Views: 10778

Re: how to increase DNA amount about 10 ug for digestion

but my protocol does not ınclude thıs buffer:s ı lyse my cells by ıncubatıng them sds and proteınase K.
by biolog35
Sat Jun 28, 2008 2:49 pm
Forum: Molecular Biology
Topic: DNA Replication-Polymerase/Primer
Replies: 3
Views: 4686

Re: DNA Replication-Polymerase/Primer

hi elena.DNA POLYMERASES NEEDS TO A TEMPLATE TO BİND BEFORE PROCEEDİNG İT İS RELATED TO ITS STRUCTURE.I DONT KNOW WHETHER U KNOW OR NOT STRUCTURE OF DNA POLYMERASE İT IS REALLY COMPLEX.IT NEEDS FREE 3' OH GROUP TO ADD NEW dNTPs SO IT NEEDS A PRIMER.AS FAR AS I REMEMBER IT HAS THREE OR MORE DOMAINS T...
by biolog35
Sat Jun 28, 2008 2:44 pm
Forum: Molecular Biology
Topic: how to increase DNA amount about 10 ug for digestion
Replies: 12
Views: 10778

Re: how to increase DNA amount about 10 ug for digestion

ı trıed ıt fruitfly but it did not work:(
by biolog35
Sun Jun 22, 2008 1:51 pm
Forum: Molecular Biology
Topic: how to increase DNA amount about 10 ug for digestion
Replies: 12
Views: 10778

Re: how to increase DNA amount about 10 ug for digestion

thanks canalon :D ı ll try tomorrow all things we discuss see u
by biolog35
Sat Jun 21, 2008 12:22 pm
Forum: Molecular Biology
Topic: how to increase DNA amount about 10 ug for digestion
Replies: 12
Views: 10778

Re: how to increase DNA amount about 10 ug for digestion

thanks canolon:D but CsCl METHOD İS efficient method ı know but it takes more time and requires large amount of reagents.it is a second solution for me now .yes u are rıght pseudomonas is diffucult to lyse and also produce large amounts of polysaccharides so CTAB method is suitable for me.also ı'll ...
by biolog35
Fri Jun 20, 2008 12:46 pm
Forum: Molecular Biology
Topic: how to increase DNA amount about 10 ug for digestion
Replies: 12
Views: 10778

Re: how to increase DNA amount about 10 ug for digestion

ok but ıt ıs too diffucult u know.ı thınk there must be another method.thaks:D