Michael David Hoos

10 Questions 11 Answers 0 Followers

Questions related from Michael David Hoos

Does anyone know of an antibody that is specific for the GOLLI region of GOLLI-MBP proteins?

I need to distinguish GOLLI proteins from their related myelin basic protein isoforms. It seems that most GOLLI-MBP antibodies are specific for the MBP containing regions of the proteins and...

12 December 2015 6,511 0 View

What are effective inhibitors of methylation for use in cell culture?

I am selecting for stable transformants in an N2a cell line. The construct looks like this: EF1a-gene of interest-IRES-eGFP...SV40-EM7-blasticidin resistance. I seem to be getting good...

09 September 2014 6,238 4 View

How can I delete a short (42bp) section of a plasmid (~9kb) and replace it with another (57bp) fragment without using restriction sites?

There are a lot of techniques that may accomplish this, such as overlap extension PCR and adaptions of Gibson, CPEC, etc. My experience with these is limited, however, and I would appreciate any...

01 January 2014 9,252 0 View

Has anyone ever experienced warping/melting of gels in a biorad mini-protean gel box?

I am running hand cast 12% bis-tris gels using MES buffer (novex formulations) at 100v. Every so often, towards the end of the run I notice a warping of the gel at the bottom corners which...

11 November 2013 9,693 1 View

Is it necessary to increase the annealing temperature after a few rounds of PCR when introducing restriction sites?

I'm designing specific primers to clone a small (~298 bp) fragment from a plasmid and introduce restriction sites. Both forward and reverse complement primers are 27 bp, no hairpins, non-self...

08 August 2013 6,987 10 View

What is the best procedure for getting RNA from small numbers of cells (<100,000)?

We are doing a cell sorting experiment and many of the fractions I would like to analyze are yielding between 50,000 - 100,000 cells. I am looking for a good procedure for high RNA yields that...

06 June 2013 2,413 28 View

Is there a good method for isolating mRNAs from p bodies? Can I simply use an Ago 2 pulldown and then isolate the mRNA from that?

I would like to know if my particular mRNA is being transitioned in and out of p bodies under certain conditions. Looking by qPCR I can see clear changes in transcript levels, but I never see...

06 June 2013 9,232 2 View

For qPCR, what should be used as the comparator?

I need to settle a disagreement with a colleague about how best to present some qPCR data. We have two mice strains (a wild-type and a transgenic), both were treated with a drug and with saline...

04 April 2013 4,062 13 View

How to do error propogation in qPCR (ddCt method)?

I have read many many papers on this and every day I come up with a different answer! My experiments usually consist of multiple transgenic animal lines including WT, treated with either saline...

04 April 2013 6,419 4 View

What is the best way to block miRNA?

miRNA seems to be at the center of translational regulation for many proteins. To study this phenomena there are many commercially available miRNA inhibitors as well as miRNA mimics. After...

03 March 2013 7,231 4 View