Hi all. We recently saw some odd Cts from a qPCR based test we use for QC'ing oligonucleotide plates. We ran tests and checked records on all of our materials and found them to be in spec. However we found our (previously in-spec) oligonucleotides to be out of spec for concentration. We assumed the cause to be within tube heterogeneity that was aggravated by aliquotting off the top layer. After broad investigation were ultimately led back to our freeze thaw prodcedures. As a check we thoroughly mixed the remaining oligos then separated them into two aliquots. We then re-ran 2 new sets of plates after thawing one set of oligos using our existing thawing protocol as well, and the other on a Box Scientific thaw station (you set plates or tubes on it and it circulates ambient air to give fast/even thawing). Interestingly enough we saw more consistent results after using the thaw station. Cts were expectedly higher but relative Cts gene to gene looked more in line with historical data. We interpreted this as lower overall variation. Does anyone have experience with identified issues in thawing proteins and oligonucleotides that led to inconsistent or unexpected data?. In the course of our investigation we were surprised to find lots of papers on the topic but didn't feel many were relevant to our exact model. But now that we have proven the point to ourselves, we are considering a wider study in house. There is little regulatory guidance around thawing itself, so we are just curious what insights the greater community may be able to offer. We are also curious if anyone has any broader experience with these thaw stations? We bought one out of curiosity but haven't used it much. However it clearly gave better results in our study, and we see now that it allows us to determine an optimum thaw time then proceduralize (is that a word? MS says no) it. Seems like better practice anyway. Anyway, just looking for thoughts on all topics. I'm new to the forum so hello and thanks for your comments.