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Questions related from Phil N Ubanako
I knocked down a gene which has been shown to promote cell proliferation. qPCR results confirmed that the gene was indeed knocked down. There was increased expression of p53 as a result, which...
09 September 2014 589 6 View
Are these 2 scenarios the same? Setting up a "No RT" control in a 2-step qPCR is done by: 1. including all components in the reaction mixture but omitting the reverse transcriptase enzyme when...
06 June 2014 3,067 4 View
My "no template" controls are showing products that are identical to their genes. I did bax and p53 qPCR and noticed that bax amplification was almost identical to bax no template control; p53,...
05 May 2014 769 8 View
I previously split confluent flasks every 48 hours. After a while (without numerous passages), the cell growth rate slowed down tremendously. In fact it has taken a month and they are yet to grow...
02 February 2014 1,084 1 View
I want to do an MTT for the first time on RMG1 and MRC5 cells. I am a bit confused about the blank and controls. Please correct me if I am wrong. Positive control=Untreated cells+MTT...
12 December 2013 2,076 26 View
I have made up 5nm,10nm and 20nm paclitaxel in culture medium and stored at 4 degrees. I have also made up 0.25, 0.5 and 0.75 micro molar camptothecin in normal culture medium, stored at 4...
12 December 2013 3,578 4 View
I wish to effectively knockdown 2 genes. I have two methods in mind: using both siRNAs at once or doing it sequentially; one knockdown after the other. I intend to proceed with flow cytometry,...
11 November 2013 5,242 15 View
I am using duplicate wells per sample in a 24 well plate. I had no problem when I extracted total RNA from a culture flask. I am using a Nucleospin extraction kit from Promega. I get low yields...
09 September 2013 1,981 23 View
I had a 700ng/ul total RNA and got 1432ng/ul of cDNA (measured using Nanodrop-Thermo Fischer). Reverse transcription was done using the Improm II reverse transcription kit (Promega). Is it...
08 August 2013 8,691 6 View
I am using a 24 well plate and the cells are too few after 24hrs and I am concerned about my RNA yield. I would need to proceed with RT PCR for gene expression studies. I am using 0.5 micromolar...
07 July 2013 9,400 9 View
Can I use RNAse-free water in the place of 'nuclease-free' water? Is RNase-free water always DNase-free OR is DNase -free water always RNase free?? The term nuclease-free water seems all...
07 July 2013 1,774 4 View
