Alisha Fleming

5 Questions 5 Answers 0 Followers

Questions related from Alisha Fleming

Expected Ct values when using TaqMan advancer mircroRNA assay?

What range of Ct values should I expect to see when using TaqMan qPCR with microRNA? What is the maximum acceptable Ct value for this kind of PCR, e.g. 30?

10 February 2017 1,409 3 View

Recommendations on how to store hiPSC cardiomyocytes before Qiagen RNA isolation?

I will be isolating total RNA from hiPSC cardiomyocytes using a Qiagen kit. Would it be better to freeze the cells straight away, then thaw and lyse, or to add lysis buffer to the cells before...

06 February 2017 6,920 3 View

Recommendations on tools to check primer alignment to gene of interest CDS?

I need to check the alignment of the primers I have designed to the target gene. Are there any online tools that don't require you to download or purchase the software as I have to request to...

17 January 2017 5,764 3 View

High ct values for GAPDH Housekeepers using cardiomyocyte cDNA compared to control cDNA?

In my qPCR reactions the housekeeper ct values for GAPDH are very high (ranging from 22-39) and Actb values are undefined when I used the cDNA extracted from cardiomyocytes. When using control...

03 January 2017 3,139 5 View

What is the minimum concentration of RNA (e.g 100ng/ul) that can be used in the applied biosystems RNA to cDNA synthesis kit?

For the RNA precipitated, most concentrations are around 10ng/ul with some even lower than 10ng/ul. The 260/230 ratios are also poor, most below 1. Can cDNA synthesis still work for these samples?

24 November 2016 6,172 6 View