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PCR for fibronectin EDA

Contributed by Dr. Alexei Gratchev

PCR for the anlysis of EDA (EIIIA) alternative splicing of fibronectin (Acc.# X02761)

Amplification of 416 bp fragment from EDA+ cDNA and 146 bp from EDA- cDNA.
Forward primer: 5'-CAA ACA GAA ATG ACT ATT GAA GGC.
Reverse primer: 5'-TGA GTG AAC TTC AGG TCA GTT GGT.


Reaction conditions
buffer 10x without Mg 2.5 µl
Mg (25mM) 1.5 µl
dNTP mix (2mM each) 2.5 µl
Primer forw (10mM) 1.0 µl
Primer rev (10mM) 1.0 µl
Taq polymerase (5U/µl) 0.2 µl
Template DNA (0.1-1µg/µl) 1.0 µl
H2O 15.5 µl

Cycling conditions for MJ Research DNA Engine
Initial denaturing 94° C 1 min
30 cycles 94° C 45 sec
60° C 1 min
72° C 1 min 30 sec
Final extension 72° C 5 min

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© Copyright 1999-2006 Alexei Gratchev. All rights reserved.