Hi there,
Just starting to make cDNA libraries and not sure how the electropherogram should look when I do fragment size analysis with Bioanalyzer High sensitivity DNA chip.
So I used the KAPA Stranded mRNA-seq kit. Fragmentation is thermal, selecting for 200-300bp fragments.
Goal is DE between drug treatments compared to control. Right now just troubleshooting to make sure our library prep is up to par before doing the real libraries.
Sample 2: 2ug total eukaryotic RNA from mouse primary neuron culture with 10x PCR cycles
Sample 4: 1ug same type of RNA with 10x PCR cycles
Sample 5: 1ug same type of RNA with 8x PCR cycles
My feeling is that Samples 2 & 4 are just over amplified/too much cDNA loaded for the high-sensitivity DNA chip.
Please give me feedback on my electropherograms, would be very greatful for any positive or negative feedback.
Cheers.
Just starting to make cDNA libraries and not sure how the electropherogram should look when I do fragment size analysis with Bioanalyzer High sensitivity DNA chip.
So I used the KAPA Stranded mRNA-seq kit. Fragmentation is thermal, selecting for 200-300bp fragments.
Goal is DE between drug treatments compared to control. Right now just troubleshooting to make sure our library prep is up to par before doing the real libraries.
Sample 2: 2ug total eukaryotic RNA from mouse primary neuron culture with 10x PCR cycles
Sample 4: 1ug same type of RNA with 10x PCR cycles
Sample 5: 1ug same type of RNA with 8x PCR cycles
My feeling is that Samples 2 & 4 are just over amplified/too much cDNA loaded for the high-sensitivity DNA chip.
Please give me feedback on my electropherograms, would be very greatful for any positive or negative feedback.
Cheers.
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