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dNTPs (0.5 無)
1 x buffer
Primer 1 (T7) (0.5 無)
Primer 2 (SP6) (0.5 無)
Taq polymerase (1 U)
Template DNA (1 無)
Top up to 25 無 with deionized H2O
PCR parameter
94C - 5mins (Initial denaturing)
*94C - 0.5min
*45C - 0.5min
*72C - 0.5min
72C - 10min (final extension)
*Repeats 30 X
Electrophoresis process (45mins at 90V)
Marker DNA 100bp (0.2 痢 ladder)
PCR product should be ~550bp
My problem
Why does my gel yields three bands? Is it something wrong with my procedures? The other two fragments are bigger than the last band. Why is it so?
I can't figure it out. Please help. Thanks in advance.
