Hosuk:LabNotesMDA/2013-2-5: Difference between revisions

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====qPCR run====


*Run qPCR with the sample from 1/28, 1/29 and 2/4 using mouse and human primer
*The number of tubes
**1/28: 16ea (#9~#24)
**1/29: 21ea (#1~#21)
**2/4: 23ea (#2~#24)
*Preparation
**1/28, 1/29 samples: 8 tubes by taking 0.5uL from two tubes and merging into one sample
**2/4 samples: 5 tubes by taking 0.5uL from four tubes and merging into one sample
*PCR master mix
**KAPA fast for 30uL reaction --> this was not used!
***KAPA fast 15 uL
***10 uM primer 1 .6 uL
***10 uM primer 2 .6 uL
***DNA from library 2 uL
***water 11.4 uL
***total 30 uL
**For 20uL reaction with mouse primer
***KAPA fast 10 uL
***10 uM primer 1 .4 uL
***10 uM primer 2 .4 uL
***DNA from library 1 uL
***water 8.2 uL
***total 20 uL
**For 20uL reaction with mouse primer
***KAPA fast 10 uL
***10 uM primer 1 .4 uL
***10 uM primer 2 .4 uL
***DNA from library 2 uL
***water 7.2 uL
***total 20 uL
PCR reaction
**95C 10sec
**58C 20sec
**72C 45sec
**40cycle
====Result====
*Most of curves rose after 24 cycle, and became flat around 32 cycle
*Seemed too much low amount of MDA product
*Human DNA has also been seen.
*(need more description)
[[File:qPCR_curve_2013-02-05.jpg|300px]]

Revision as of 17:40, 6 February 2013