User:Marcel Quint/Notebook/medicago/2008/05/28

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Medicago
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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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RT-PCR of medicago Aux/IAAs - internal standard test
35 cycles → [2] for Actin F/R (Ta=53) + [1] for ActinMt F/R, β-Act F/R, and AC138130_5 (Ta=50) primers: Actin F/R → ? bp ActinMt F/R → ? bp β-Act F/R → ? bp AC138130_5-F/R	→ 981 bp
 * use cDNAs from 4/25 and dilute 1:10 → use 2 µl of this dilution as template for PCR
 * use AC138130_5 as positive control for PCR rx
 * 25 µl DreamTaq Rx:


 * gel 1% LE:

row1

row2

results: taken together: use ActinMt F/R as internal standard with Ta=50°C, try 30 cycles
 * 1) Actin F/R: ok, very short fragment,use less cycles
 * 2) ActinMt F/R: ok, very short fragment,use less cycles
 * 3) β-Act F/R: root does not come up, discard


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