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CPRES's Content

There have been 167 items by CPRES (Search limited from 17-April 20)



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#173189 Converting clinically relevant dose for in vitro studies

Posted by CPRES on 03 February 2015 - 06:18 PM in MTT, Proliferation and Cytotoxicity Assay

Bump. I am also interested in finding out how they do it. I mean actually, not guesswork.




#173188 What is your IQ?

Posted by CPRES on 03 February 2015 - 06:16 PM in Chit Chat

Just 10.

 

cubed.




#173187 Words, Words, Words

Posted by CPRES on 03 February 2015 - 06:14 PM in Chit Chat

encephalomyeloradiculitis




#173186 is -40 freezer enough for cell freezing?

Posted by CPRES on 03 February 2015 - 06:10 PM in Cell Biology

Yes, you can transfer from -40 to LN2 without problem. I have done it several times with several cell lines, out of impatience, so I am sure it works.

 

However, don't do it with some sensitive or irreplaceable cell line without doing some trial runs. And if you do, don't sue me. What's with your Avatar name by the way?




#173185 Software to create nucleotids

Posted by CPRES on 03 February 2015 - 06:03 PM in siRNA, microRNA and RNAi

I am not sure I understand what exactly you are looking for. Can you elaborate?




#173184 U251 and U87 cells

Posted by CPRES on 03 February 2015 - 06:01 PM in Tissue and Cell Culture

I can't provide you personal advise, but you should find some information here and here.




#173183 Can I skip the FC blocking step in flow cytometry?

Posted by CPRES on 03 February 2015 - 05:55 PM in Flow Cytometry

As Tab said, however, you can do away this step if you are staining whole blood (because there is a lot of serum in the whole blood blocking the Fc receptors).




#173182 Transformation E.Coli help

Posted by CPRES on 03 February 2015 - 05:50 PM in Molecular Cloning

No. Please update us on how profound was your disappointment when you opened the incubator tomorrow :)




#173093 Gene expression and transcription factors

Posted by CPRES on 30 January 2015 - 08:20 AM in Molecular Biology

gene expression is controlled by several things:

 

1. Transcription factors (repressors and activators)

2. DNA epigenetic changes

3. Histone epigenetic changes

4. Enhancers (long-range)

5. Silencers

 

In addition, once a genomic transcript is generated, it can be spliced, truncated, degraded in so many ways that it changes eventual protein levels. Also, protein can be controlled in several ways (ribosomal processing, tagging for degradation etc) during and after translation. 

 

And this is very basic idea, there is a lifetime of complexity to talk about here.




#173092 Anyone has a experience with Fluorescently labelled siRNA ?

Posted by CPRES on 30 January 2015 - 08:13 AM in siRNA, microRNA and RNAi

No, here is what you do:

 

1. Buy a siRNA that is targeting your gene. This is unlabeled siRNA directed against your gene.

 

2. Buy a control (BlockiT) alexa fluor red siRNA. This siRNA is carefully designed such that it does not target any mammalian gene. So, it is a negative control, but more importantly, it is a transfection efficiency control.

 

And that is all you need. Forget about a kit to label your siRNA, unless you want to see its cellular localization and dynamic. Most experiments are just about transfecting siRNA to knockdown the gene of your interest.




#173089 Words, Words, Words

Posted by CPRES on 30 January 2015 - 07:52 AM in Chit Chat

La gecko




#173088 SDS page

Posted by CPRES on 30 January 2015 - 07:50 AM in SDS-PAGE and Western Blotting

Thanks for the feedback!




#173087 Anyone has a experience with Fluorescently labelled siRNA ?

Posted by CPRES on 30 January 2015 - 07:47 AM in siRNA, microRNA and RNAi

The BLOCK-iT Alexa Fluor Red is already labeled siRNA, so you don't have to label or incorporate anything (if that is what you are concerned about).

 

Just use this fluorescent siRNA exactly as you use your experimental siRNA in the protocol.




#173080 Is it possible to "over-block" a membrane?

Posted by CPRES on 30 January 2015 - 06:26 AM in SDS-PAGE and Western Blotting

Wine is more like women, not all improve with age, but some do. Whiskey is more like men, all of them mellow with age.




#173078 Anyone has a experience with Fluorescently labelled siRNA ?

Posted by CPRES on 30 January 2015 - 06:20 AM in siRNA, microRNA and RNAi

There is nothing to it. You will be able to see if you are getting a good transfection efficiency with your method.

 

Basically, you transfect your experimental siRNA and control (fl labeled) siRNA to your cells (in separate wells of course). Then check the control cells under fl microscope to see what % of cells show fl. 




#173077 Words, Words, Words

Posted by CPRES on 30 January 2015 - 06:15 AM in Chit Chat

el capitan




#173076 Is it possible to "over-block" a membrane?

Posted by CPRES on 30 January 2015 - 06:11 AM in SDS-PAGE and Western Blotting

Wine, Whiskey and Western block, all improve with time.




#173048 Slight Dilemma

Posted by CPRES on 29 January 2015 - 07:33 AM in Dissertation and Paper Writing

Labnerd: "However, I feel like if we submit the manuscript to this journal, it will bypass the editorial board and head straight to the reviewers, which I think will pretty much guarantee an acceptance."

 

This is precisely you should not submit to the journal. Just as you think you can bypass the process, other people will think you probably bypassed the process and paper was otherwise not much worth. Go with your supervisor's advice. Believe in your work.




#173047 Grants for research

Posted by CPRES on 29 January 2015 - 07:31 AM in Life as a PhD Student and Postdoc

Depends upon what level of training you have. BS, MS, PhD? You can come over to US for a fellowship if you are already PhD.




#173046 re-staining after IHC of mouse brain slices

Posted by CPRES on 29 January 2015 - 07:29 AM in Histology and Pathology

Hi Funny Fish, I don't want to discourage you, but I have attempted such thing and has never worked for me. But who knows it might work for you.

 

If you have enough unstained sections with you, it is always best to do a clean experiment. 




#173045 Unidentified structure in tongue

Posted by CPRES on 29 January 2015 - 07:26 AM in -Anatomy, Physiology and Zoology-

You got to do some work, man!

 

It took me 30 seconds to view your photograph, and 30 seconds to do google image search (q=tongue histology), to find the answer.

 

Do it. Then if you still have question, give us what answer you came up with and I will tell you my answer.

 

Edit: You are lucky to be living in Google age, otherwise you would have to go pick up a histology atlas. And that is what you should do anyways.




#173006 No colonies on plate after liagtion

Posted by CPRES on 28 January 2015 - 04:40 AM in Molecular Cloning

There are so many things to point out, so I wouldn't. But as you say " i did have very low dna concentrations of vector n insert", I think that could be the reason. YOu should at least have 50-100 ng of linearized, purified plasmid in the ligation reaction. That is crucial. Then comes the 1:3-1:5 molecular ratio of vector:insert. So in your case you need 100 ng vector and 25 ng of insert in your reaction. Or 50+12.5. But don't go below 50 ng for linearized vector.




#173005 Estimate cost for Molecular Docking software/devices

Posted by CPRES on 28 January 2015 - 04:32 AM in Molecular Biology

Just call up or email the companies, they will come knocking on your door! 




#173004 Question about MBP purification

Posted by CPRES on 28 January 2015 - 04:29 AM in Protein Expression and Purification

Here is a big list of available protein binding resins.




#173003 A problem with PCR array

Posted by CPRES on 28 January 2015 - 04:23 AM in PCR, RT-PCR and Real-Time PCR

This is too general a question. There could be one or two of about 74 things that could be going wrong. See here for some PCR array manuals and troubleshooting guides, think critically, get help from a senior colleague to get an objective assessment of your techniques that you may be missing.





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