Sequencing Lambda DNA

Rene.DeMot at Rene.DeMot at
Tue Mar 28 06:11:08 EST 1995

We have sequenced a part of the insert of a lambda clone (EMBL3) after 
subcloning in pUC19. Now we are trying to sequence into the flanking 
sequences using a primer based on the sequence already known.
In order to avoid subcloning, we are looking for a protocol suitable for 
sequencing lambda DNA directly. We have tried some denaturation protocols and 
varied the amounts of DNA, but with little success. The sequencing primers 
work well when using them on pUC-cloned DNA.
Could somebody provide a protocol suitable for the A.L.F. sequencer (using 
fluorescently-labeled primers)?

Rene De Mot

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