Kpn I site blunting

Hirata, Masakazu hiratamz at MFOUR.MED.KYOTO-U.AC.JP
Mon Nov 20 00:38:13 EST 1995


Hello!
 I have tried to blunt a Kpn l site by Klenow fragment and ligate a 
PCR fragment into the blunted site.
 When I sequenced the ligation part, the site contained 
	 GGxxxx (xxxx being PCR primer sequence) instead of Gxxxx.
 Since Kpn I site is GGTAC|C, 
I expected it to become single G at its 3' end after blunting with  
Kpn I.
 However, that is not the case. 
 I have sequenced seven independent clones and got the same result.
 Does this mean Kpn I site is actually GGTA|CC?
 This may be a FAQ, but I could not find any bionet document from 
bionet gopher search engine.
Thanks in advance for any information.

Masakazu Hirata
Kyoto University Faculty of Medicine



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