Nathan Johnston

4 Questions 10 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...

30 January 2014 9,444 13 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...

22 October 2013 747 12 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...

13 June 2013 4,602 9 View

Ficoll vs Percoll (discontinuous vs continous) in isolation of TILs from melanoma?

I am trying to isolate tumor infiltrating lymphocytes from melanoma tumors (B16F10) injected into mice subq and am having three issues with what is the best way to go about it. In the end I need...

08 May 2013 9,320 9 View