Gurudatta B V datta at ccmb.res.in
Mon Jun 9 09:43:32 EST 2003

Dear all the FLY GURUs
I Have some doubts in my mind I ope there will be some solution with you
kindly reply at datta at ccmb.res.in
1.If you are desiging the ds RNA construct in 3`UTR .Does this have the
Trnascription termination signal (Aauaaa)!!. or this does not have any
role and express the Reverse tandem !!

2.When i have put first target sequnce and While putting second sequnce
after 19 bp Polylinker in Xba I site as reverse tandem .I am not finding a
single clone Even after seceeng 120 colonies(THe xba is treated With SAP
And back grone is mimised)

HAs any body faced this problem !! IS it due to repeat sequnce There is
no ligation .
There is recombination happening ind"DH5 alpha rec - stain 

please suddest some  thing 

thanking you 


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