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Old 09-23-2016, 09:32 AM   #6
Conradona
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Location: Berlin, Germany

Join Date: Aug 2016
Posts: 2
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Dear all,

I am preparing single-cell libraries according to the method described in Macaulay I, et al. G&T-seq: Parallel sequencing of single-cell genomes and transcriptomes. Nature Methods, 2015.

The amplified cDNA seems to look fine on the tapestation when I add no ERCC spike-in controls.

But I get strange double peaks when I add the controls and the tapestation screentape gets overloaded. The paper says to use 1 μl of a 1:250,000 dilution and I went up to 1:500,000 and 1:1,000,000 but get the same results.

The project leader tells me to just dilute more and more but I am suspicous that this won’t solve the problem.

Thanks in advance for any hints!
Conny
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File Type: jpeg no ERCC.jpeg (125.6 KB, 62 views)
File Type: jpeg with ERCC.jpeg (136.0 KB, 48 views)
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