Hello,
I recently generated cDNA from 100 T-cells (from the same source) using the Smart-Seq2 protocol from Sandberg et al. After the pre-amplification step (all wells were pre-amplified for 18 cycles), I ran 1uL of my cDNA on a Bioanalyzer and noticed that the amount of cDNA that I recovered was relatively variable (range from ~450pg/uL to ~1,400pg/uL).
Does anyone know how much variance in cDNA concentration is expected well-to-well? Is this variance biological or an artifact of the protocol?
I would like to get an idea I am worried that the variance that I observe is due to technical error on my part, perhaps in pipetting.
Thanks in advance for the help!
I recently generated cDNA from 100 T-cells (from the same source) using the Smart-Seq2 protocol from Sandberg et al. After the pre-amplification step (all wells were pre-amplified for 18 cycles), I ran 1uL of my cDNA on a Bioanalyzer and noticed that the amount of cDNA that I recovered was relatively variable (range from ~450pg/uL to ~1,400pg/uL).
Does anyone know how much variance in cDNA concentration is expected well-to-well? Is this variance biological or an artifact of the protocol?
I would like to get an idea I am worried that the variance that I observe is due to technical error on my part, perhaps in pipetting.
Thanks in advance for the help!
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