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文献和实验contamination in real-time pcr negative control-Real-Tim
carryover from the pipet, or primer dimer? Thanks!!! -medhan1012- I had this trouble too, and sometimes still happens but after 35-40 cycles. If You get a signal in negative control under the 35-30 cycles You're having troubles. If You run
: mycoplasma DNA ( A. laidlawii ; M. pirum ) 3.1.1.3 Negative control : ddH2O 3.1.2 Reaction mixture ( 23 ul / each ) 3.1.2.1 10x PCR buffer (含1.5 mM MgCl2 ) 2.5 ul 3.1.2.2 1st stage primer mixture 0.5 ul 3.1.2.3 dNTP ( 1.25 mM each ) 1.0 l 3.1.2.4 MgCl
24 µl H2O and 1 µl RNase inhibitor, 20 U/µl, to RNA pellet. Resuspend the RNA pellet with gentle tapping. Quick spin. Aliquot 12 µl into 2 tubes for the (+) and (-) RT reactions. Prepare additional control tubes using the following in place
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