Hello
I picked 100, 150, 200 neuronal cells from mouse brain tissue using arcturus lcm.
extracted RNA using picopure kit and then using SMARTer Stranded Total RNA-Seq Kit - Pico Input Mammalia kit I made libraries.
The bioanalyser traces (appended along) show spiky peaks ranging from 300bp to 23000bp. The spikes are beyond the 10380bp markers (shown in purple). I have following questions:
1.Are these spikes PCR artifacts (I took 16 cycles of PCR).
2.Can I do gel extraction from these libraries from size ranging from 300 to 1000bp, if yes, then what is the best method?
For reference:
sample 1,4,7 and 10 are from RNA extracted out of 100 cells
sample 2,5,8 and 11 are from the RNA extracted out of 150 cells
and samples 3, 6 and 9 are from 200 cells.
Thanks in advance,
Ram
I picked 100, 150, 200 neuronal cells from mouse brain tissue using arcturus lcm.
extracted RNA using picopure kit and then using SMARTer Stranded Total RNA-Seq Kit - Pico Input Mammalia kit I made libraries.
The bioanalyser traces (appended along) show spiky peaks ranging from 300bp to 23000bp. The spikes are beyond the 10380bp markers (shown in purple). I have following questions:
1.Are these spikes PCR artifacts (I took 16 cycles of PCR).
2.Can I do gel extraction from these libraries from size ranging from 300 to 1000bp, if yes, then what is the best method?
For reference:
sample 1,4,7 and 10 are from RNA extracted out of 100 cells
sample 2,5,8 and 11 are from the RNA extracted out of 150 cells
and samples 3, 6 and 9 are from 200 cells.
Thanks in advance,
Ram
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