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Yeah, we were quite surprised when I spoke to them and discovered this. In the future we might ask them to use all and increase the cycles to 14 or 15. That would explain A LOT of reasons why we are struggling with our samples (RNA, not so much for DNA).
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Hi jteeee2,
I just wanted to check back in with you and say thank you. We did some testing in the lab and both the primers / Taq you recommended appeared to work well. We submitted that to our Sequencing CORE and we were able to save over 2/3's of those 68 samples. That a huge relief!!! So instead of all 4 out of 72, we now have material for 49. We have a plan to save those other 23 samples and we are asking them to increase the cycles from 12 for future RNA/FFPE runs.
Thank you so much.
SueComment
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Glad to hear things are improving. Best of luck getting those last 23 samples made into libraries.Comment
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Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.07-01-2026, 11:43 AM
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