Attached I have a D5000 tapestation from a partially prepared library from a new protocol i'm trying. This is a pooled library prep where cells are lysed a 384-well plate in 1% triton and RT mastermix (containing primers that have unique barcodes to identify the material from each well) is added directly to each well. After RT, the well contents from the plate are pooled into one tube, the cDNA is purified and then amplified for 26 cycles. This tapestation shows an anlysis of the resulting material. I'm curious about the unique shape of the product (around 10,000 average size). Has anyone seen a tapestation result that looks similar to this one before? I used this material to run a qPCR on a panel of housekeeping genes (GAPDH, ACTB, etc.) that should be expressed in my cell type but I didn't get any amplification. I'm wondering what this product could be and if I should just deep sequence it to see whether my pooled library preparation method is working?
- Badges
- Science topic
- Similar topics
- Biological Science
- Histology
- Sectioning