IUBio

library construction problem

John Ladasky jladasky at pmgm.Stanford.EDU
Wed Mar 10 07:31:54 EST 1999


Frederik,

	Which lambda phage vector are you using?  The lambda phage
genome needs to be within a certain length range in order to be 
packaged.  Some vectors, like Lambda Zap, will take inserts of 
negligible size, up to 12 kB.  The Lambda Fix vector, on the other
hand, takes inserts of 9-23 kB.  If you are using Lambda Fix with
inserts under 10 kB, most of your phage genomes will be too short.

In article <36E6B664.DD4 at nospam.ipk-gatersleben.de>,
Frederik Boernke <boernke at nospam.ipk-gatersleben.de> wrote:
[snip]
>fraction containing the desired fragment size was resolved on an agarose
>gel and fragments ranging from about 3 to 10 kb were excised and
>purified using Qiaex gel purification kit (Qiagen). 2.5 ul purified DNA
>were ligated into BamHI/CIAP treated lambda arms (Stratagene) in a total
>volume of
[snip]

-- 
Rainforest laid low.
"Wake up and smell the ozone,"
Says man with chainsaw.					- John Ladasky



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