I have 3 inserts one is 464 bp, 1173 bp and 713 bp. I am trying to ligate and amplify them using pcr. I have done a overnight ligation and found very faint bands on my gel. I try to amplify though nothing is being amplified.
I have been using this protocol to extract dna from fission yeast though it is not working. I am getting two bands that are very small in size. Can someone troubleshoot with me...
07 August 2018 1,890 0 View
I am wanting to fuse 3 dna together and insert them in a plasmid. Is fusion pcr?
11 December 2017 1,584 0 View
I am looking for a protocol using 3 dna pcr.
11 December 2017 4,511 4 View
I am trying to fuze 3 different inserts together. I am looking for a good fusion protocol.
11 December 2017 3,583 2 View
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
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
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
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
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
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
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
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
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
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