Martin Akandawen

7 Questions 11 Answers 0 Followers

Questions related from Martin Akandawen

Why do I see amplification in my negative control?

I am running RT-qPCR for a gene. However I still see amplification with lower Cq values in my negative control. Previously the Cq values of the negative controls were above 30. But now it averages...

04 July 2023 7,184 4 View

Can DNase simply be added to extracted RNA solution, incubate at RT and enzyme heat killed??

I extracted RNA from single cells using RNAeasy kit. I did add DNase I as stated in the extraction protocol. However my negative control (without reverse transcriptase) still showed amplification...

17 April 2023 6,620 5 View

Should RNA or cDNA or both be normalized for qRT-PCR and why?

I am quantifying level of gene expression using qRT-PCR. I want to know at what stage is best for normalization of samples. Is it at the Reverse transcription stage (RNA normalization) or PCR...

07 April 2023 6,671 8 View

Does freezing mouse spleen in liquid nitrogen to inactivate RNase affect CD8 T cell isolation?

I want to harvest RNA from CD8 T cells from mice spleens. In order to minimize RNA degradation I intend to freeze the spleens in liquid nitrogen before homogenization. Will this affect CD8 T cell...

09 January 2023 4,635 2 View

HPLC: How do you determine the expected concentration of analyte to inform you of the amount of internal standard to add to sample for analysis?

The amount of internal standard to be added to the sample for analysis is suppose to be about 0.3 to 0.5 of the expected amount of the analyte. How do you determine the expected amount of the...

23 March 2018 2,461 4 View

Which is the better administration for effect of an extract on PK of a drug studies?

I want to know which of the following is the better administration for PK studies and why 1. Administration of extract alone for 13 days. Overnight fast and administration of drug for PK...

22 March 2018 3,821 1 View

How does internal standard ensure accuracy by catering for sample losses through flow rate and injection volume in HPLC analysis?

I am looking for explanation on how internal standard in HLPC analysis help in obtaining accurate results in the event of sample losses in the course of injection and flow rate processes

22 March 2018 3,686 4 View