Which company has the best BamHI and EcoRI?

Hello Everyone, I am curious to know your experiencers from different brands for the above enzymes. Also if these are a good restriction enzyme combinations for double ligation of 3 PCR products...

31 December 2017 4,423 9 View

Does anyone use saturated trehalose (~75% W/V) for reverse transcription or PCR?

How do you make and store saturated trehalose? What do you do to dissolve it? '. Thank you!

03 April 2015 606 1 View

Could anyone explain how the coverage is calculated in amplcon seq?

31 December 2014 3,756 2 View

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

Alginate and silk fibroin hydrogels toxic to cells?

Hello, I asm currently working with alginate and silk fibroin hydrogels. I prepared a blend of alginate and aqueous silk fibroin (straight from dialysis) and resorted to ionic crosslinking to...

23 February 2021 8,249 2 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

Why the nucleic acid gel melted after running for 30min?

Hi all, I proceed with nucleic acid gel electrophoresis recently several times, but a wried phenomenon is they always melted after running over 30min. My gel concentration is 1% percentage...

22 February 2021 9,896 3 View

Sanger Sequencing results not matching experimental results of Restriction Digestion, colony PCR and protein expression?

I have cloned two domains of a gene into two bacterial vectors using cut and paste cloning with different RE sites. I checked the putative clones by colony PCR and Restriction Digestion (fallouts...

18 February 2021 6,736 4 View