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

08 September 2014 6,198 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...

31 December 2013 9,191 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...

10 November 2013 9,654 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...

07 August 2013 6,892 10 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...

05 June 2013 9,198 2 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...

05 June 2013 2,372 28 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...

03 April 2013 6,341 4 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...

03 April 2013 4,021 13 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...

02 March 2013 7,189 4 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

How to calculate BREQ 3 questionnaire?

how to calculate BEHAVIOURAL REGULATION IN EXERCISE QUESTIONNAIRE BREQ 3 ,which contain 24 questions and each question has 5 answers (0- Strongly disagree , 1-disagree, 2- Neither agree nor...

02 March 2021 811 3 View

I am interested in medicinal chemistry field, which books are highly recommended to learn from?

I am at the beginner level in drug design and drug discovery, and I am interested to learn more about this field. Which books are highly recommended to start from and gives me all essential...

02 March 2021 8,437 2 View

Please suggest a good text book for "Research Methodology in Life Sciences". Which book would be good for PhD Scholars?

Please provide Book Title and author name

02 March 2021 9,059 3 View

Could any one guide me, to write research proposal for grant? ?

We have one plant, the local people using this for controlling blood sugar, the toxicity and antidiabetic activity of the plant is not reported yet, please guide from whag should be the process ?...

02 March 2021 2,300 1 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

Is anyone familiar with the cyclic compressive fatigue testing (BS EN 12697 - 25) ?

I would like to get more information on the cyclic compressive fatigue testing (BS EN 12697 - 25) which is typically done for bituminous material in pavement design. Any one can advise on the...

28 February 2021 2,004 3 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

What is Flat Lens Antenna?

28 February 2021 5,433 3 View

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View