User:Marcel Quint/Notebook/medicago/2008/04/29

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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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repeat (from 4/28) RT-PCR of medicago Aux/IAAs
35 cycles → [1] for AC138130_5 and AC152423_24 (Ta=50) + [2] for AC175828_16 and AC146866_11 (Ta=53) primers: AC138130_5-F/R	→ 981 bp AC175828_16-F/R	→ 810 bp AC146866_11-F/R	→ 1044 bp AC152423_24-F/R	→ 627 bp
 * use cDNAs from 4/25 and dilute 1:10 → use 2 µl of this dilution as template for PCR
 * 20 µl ExTaq Rx:


 * gel 1% LE:

row1

row2

results: from yesterday:
 * 1) AC138130_5: ok, but increase Ta to 53°C + use less cycles
 * 2) AC152423_24: fine → final conditions: 35 cycles, Ta=50°C
 * 3) AC175826_16: fine → final conditions: 35 cycles, Ta=53°C
 * 4) AC146866_11: better, but still additional bands → increase Ta to 55 and 57°C
 * 1) AC148405_5: make new primers
 * 2) AC158504_5: Ta=50°C ok, but use less cycles → try 32 and 29


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