Nathan Johnston

4 Questions 13 Answers 0 Followers

Questions related from Nathan Johnston

How do I introduce a 4 nt mutation in DNA?

I am studying how well various miRNAs bind to and silence my 3' UTR of interest. I ordered a dual luciferase plasmid from promega and now need to amplify my 3' UTR of interest to insert into the...

01 January 2014 4,703 14 View

How long should I make a 3' UTR to test miRNA silencing?

I am going to use this plasmid http://www.promega.ca/products/reporter-assays-and-transfection/reporter-vectors-and-cell-lines/pmirglo-dual-luciferase-mirna-target-expression-vector/ to test...

01 January 2014 7,356 34 View

Where to find the forward primer sequences for various miRNAs when analyzing miRNA expression using RT-PCR?

Does anyone know how to find or create the sequence for the forward primers used when confirming microRNA levels with Reverse Transcriptase-PCR? My supervisor wants me to try to find them so we...

10 October 2013 1,011 14 View

Isolating CD3+ T cells from mouse tumor tissue (C57LB/6 and BALB/c) - MACS Beads

I am injecting B6 and BALB/c mice with B16F10 and 4T1 cells, respectively, to generate melanoma and breast cancer models. I then want to extract the tumors, digest in Collagenase IV/DNase I and...

06 June 2013 4,566 10 View