Ligation of PCR products

Andreas Pahl macbeth at cs.tu-berlin.de
Thu Jul 25 06:18:16 EST 1991


In article <1991Jul19.131027.10619 at nrcnet0.nrc.ca>, num208jn at MBDS.NRC.CA (John Nash) writes:
> In article <9107182217.AA23884 at genbank.bio.net>, ST403161 at BROWNVM.BROWN.EDU (Michael Newstein) writes:
> >I have read that it is exceedingly difficult to blunt-end ligate products from
> > PCR  (without cleaving first in internal restriction sites). I am attempting
> >to clone the product of a PCR reaction into a blunt end cutter site (eg SMA 1)
> >in pBluescript. Does anyone have suggestions?
> 
> ...
 
> By the way, when I cloned my 650 bp PCR product, the positive clones
> were all light blue.... I was cursing because the transformation
> plates had few white colonies.
> 
> Hope this helps.
> 

Did you isolate the DNA from all the colonies?? How did you confirm the
positive clones??

Bye, Andreas

-- 
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Andreas Pahl                                email: macbeth at opal.cs.tu-berlin.de
Institut f. Biochemie u. Molekulare Biologie       macbeth%opal at DB0TUI11.BITNET
TU Berlin                                          macbeth%opal at DB0TUI11.EARN
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D-1000 Berlin 10                                          Fax  +49 30 314 24783
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