User:Marcel Quint/Notebook/medicago/2008/05/29

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


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RT-PCR of medicago Aux/IAAs - internal standard test

 * use cDNAs from 4/25 and dilute 1:10 → use 2 µl of this dilution as template for PCR
 * repeat reaction from yesterday, but with less cycles
 * 25 µl DreamTaq Rx:

28/31 cycles → [1] for ActinMt F/R (Ta=50) primers: ActinMt F/R → ? bp

row1 {| !lane !sample
 * gel 2% 3:1:
 * 1
 * ActinMt F/R 28 cycles - petiole2
 * 2
 * ActinMt F/R 28 cycles - stem2
 * 3
 * ActinMt F/R 28 cycles - roots4
 * 4
 * ActinMt F/R 28 cycles - leaves2
 * 5
 * ActinMt F/R 28 cycles - flowers1
 * 6
 * ActinMt F/R 31 cycles - petiole2
 * 7
 * ActinMt F/R 31 cycles - stem2
 * 8
 * ActinMt F/R 31 cycles - roots4
 * 9
 * ActinMt F/R 31 cycles - leaves2
 * 10
 * ActinMt F/R 31 cycles - flowers1
 * 11
 * pUC19 MspI marker
 * 8
 * ActinMt F/R 31 cycles - roots4
 * 9
 * ActinMt F/R 31 cycles - leaves2
 * 10
 * ActinMt F/R 31 cycles - flowers1
 * 11
 * pUC19 MspI marker
 * 11
 * pUC19 MspI marker
 * pUC19 MspI marker

results:
 * looks fine, but root sample seems to contain less cDNA
 * repeat root tissue and two other tissues for comparison with increasing amounts of root cDNA: 2-4-6-10 μL template
 * 29 cycles and Ta=50°C

row2
 * gel 2% 3:1: