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experiments related to epigenetics

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#1 DeniseB

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Posted 17 May 2013 - 05:09 AM

I wanted to take a minute and say I am new to trying anything with Epigenetics, other than reading about the field and reading research papers.
At the moment I have a BS degree and would like to perhaps pursue more education in the future studying Epigenetics, specifically gene expression resulting in chronic low dose exposure to environmental elements. Its been an interest for years, after hanging out with many chemophobic individuals, and studying toxicology while pursuing my degree.
However, I have NO LAB SKILLS. I have read more protocols than I can count but have no idea where to get started. I was going to start with simple yeast and perhaps get my hands on some that has been modified and go from there. However other than culture techniques and extractions I am clueless after that point.
If you guys have any general experiments I can run to gain some skills please share them.

Thanks.

#2 Curtis

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Posted 17 May 2013 - 10:15 PM

Welcome to the forum. We usually write our question in the title. I just edited it.

Just don't worry. Most people spend the first few months preparing reagents and read articles. You must do the same...since you want to work on epigenetics then you'll need to learn PCR and electrophoresis. Do come back if you have more questions.

#3 DeniseB

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Posted 18 May 2013 - 06:49 AM

Thanks. Funny how after I asked this question yesterday I spent the rest of the morning reading up on PCR and Electrophoresis. I will certainly be back with many questions.

Once again Thanks.

#4 et2b

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Posted 25 August 2013 - 11:22 PM

Hi Denise,

 

Reading your interests you will most likely be doing studies in human populations or in animal models.

I would start with the basics: my students usually do.

 

Isolating DNA :  using kits (Qiagen is basic supplyer) and the old fashioned way: 25:24:1 and ethanol precipitation.

Performing RNA isolations: best is to use a kit often.

Perhaps you have somebody in your center that can tap some blood? you will need some to practice!

 

Performing bisulfite reactions: when you have your first DNA use a bisulfite conversion kit to convert your DNA for DNA methylation measurements

Perform your first BS-PCR, bisulfite PCR.  Look in literature to the most convincing example of a promoter affected by a chemical and start with repeating this measurement and see if your 'control' DNAs have the same level as their control DNAs

 

Do a taq man assay and get yourself familiar with q-PCR for gene expression measurements: read up on the delta delta Ct method (most students find this excidingly complicated, so ask and get help!)

Any left over taq-man reagent will do of a friendly lab (its expensive to buy), just get your fingers going and get some measurements done before you start your own work. 

 

good luck and enjoy.






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