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Pangea's Content

There have been 97 items by Pangea (Search limited from 16-April 20)



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#147637 Restriction Enzymes

Posted by Pangea on 07 January 2013 - 01:26 PM in Molecular Biology

Is it possible to get the genes of the general used Restriction Enzymes from NCBI?



#147622 Avoiding PTM Actylation of rProtein

Posted by Pangea on 07 January 2013 - 12:28 PM in Protein and Proteomics

No of course inhibition will be happing on the protein level. I mean cutting out the gene will be lethal. But thanks for sharing your knowledge.



#147604 PTM in E. coli

Posted by Pangea on 07 January 2013 - 10:44 AM in Protein and Proteomics

Which posttranslational Modification has E.coli.?



#147601 Avoiding PTM Actylation of rProtein

Posted by Pangea on 07 January 2013 - 10:09 AM in Protein and Proteomics

Is there a way to screen a inhibitor collection?



#147600 Avoiding PTM Actylation of rProtein

Posted by Pangea on 07 January 2013 - 10:09 AM in Protein and Proteomics

This option also came in my mind but to knockout the gene would be Lethal i guess. In mammalian maybe one could use stable cell lines transfected with shRNA, or?



#147594 Replication of the Plasmid DNA

Posted by Pangea on 07 January 2013 - 07:33 AM in Genetics and Genomics

Thats shouldnt be a big deal, isnt?



#147593 my competent cell grow on any agar plate

Posted by Pangea on 07 January 2013 - 07:32 AM in Microbiology

Under which condition are you growing the stock solution cells before transforming with plasmid to have enought cells? Or are you using a stock aliqout?



#147561 cloning: band at different site than desired after RE digestion

Posted by Pangea on 06 January 2013 - 01:07 PM in Molecular Cloning

You have right you have single cut RE. Sorry about confusing.



#147549 Large-Scale Purification of Non-Secreted Protein in E.Coli or CHO Cells

Posted by Pangea on 05 January 2013 - 03:38 PM in Protein Expression and Purification

How can you lyse the cells if you have 1000 l or more? And purify from a Bioreactor?



#147548 Replication of the Plasmid DNA

Posted by Pangea on 05 January 2013 - 03:25 PM in Genetics and Genomics

You mean the Replication machinary.



#147532 Insulin production

Posted by Pangea on 05 January 2013 - 12:59 PM in Protein Expression and Purification

What would be the best way to express and purify active insulin?



#147518 Ethidiumbromide

Posted by Pangea on 05 January 2013 - 07:35 AM in Molecular Cloning

Thanks i will keep in mind.



#147517 Mycoplasma Test Kit

Posted by Pangea on 05 January 2013 - 07:23 AM in Tissue and Cell Culture

What is the best Mycoplasma Kit? Some of them are always positive or negative.



#147515 Ethidiumbromide

Posted by Pangea on 05 January 2013 - 07:10 AM in Molecular Cloning

Can someone suggest a alternative to EthBr?



#147513 High Yield Protein Expression 10g/L

Posted by Pangea on 05 January 2013 - 06:47 AM in Protein Expression and Purification

It will be a pET vectorsystem for E coli. One can play with media and ZnCl2, Glucose and VPA. But is that enought for very high yield production.?



#147511 Questions about proper technical approach

Posted by Pangea on 05 January 2013 - 05:32 AM in Molecular Biology

Julio sounds good. Something new again. Thanks for sharing you knowledge.



#147510 Spin Column

Posted by Pangea on 05 January 2013 - 05:21 AM in General Lab Techniques

Cheers very helpful.



#147502 Spin Column

Posted by Pangea on 05 January 2013 - 02:18 AM in General Lab Techniques

Where can i get single spin columns for miniprep?



#147501 TRYPSIN

Posted by Pangea on 05 January 2013 - 02:13 AM in Molecular Cloning

Thanks for your Advice.



#147500 Changing Genomic Regions in Host Cell

Posted by Pangea on 05 January 2013 - 01:46 AM in Genetics and Genomics

How can i best delete a gene region of my host cell.? And how long time will it take?



#147498 Questions about proper technical approach

Posted by Pangea on 05 January 2013 - 01:32 AM in Molecular Biology

Maybe you can check first whether it is a gene or not by using biotool like finding ORF, start and stop codons etc. Than you should clone this region if possible with this promotor.



#147497 cloning: band at different site than desired after RE digestion

Posted by Pangea on 05 January 2013 - 01:18 AM in Molecular Cloning

It might be that your digestion of your Insert is not working out you have blunt ends. So it will just ligated several inserts. But i am not sure. And i hope you are not using BL21 for plasmid replication. Not recA mutation. But BLR have it or other K 12 strains. And are sure that you are not cutting into your plasmid?



#147496 protein concentration and Pull down

Posted by Pangea on 05 January 2013 - 01:10 AM in Biochemistry

No, of course if you use more magnetic beats you will increase you yield depending on your experiment design. But as I said you beads have capacity ana useing a lot of beats are not recommend. But in different concentration as you said,



#147495 Replication of the Plasmid DNA

Posted by Pangea on 05 January 2013 - 01:01 AM in Genetics and Genomics

My Question would be which enzyme do we need in replicating Plasmids in E.coli? Any idea.



#147494 siRNA in E.Coli

Posted by Pangea on 05 January 2013 - 12:57 AM in siRNA, microRNA and RNAi

Sounds good.Thanks.




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