Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a... | Contact experts in Plasmids to get answers
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Questions related to Plasmids
I run some plasmid samples with 1% agarose gel (like the picture shown), and the band is not clear, I use 5ul DNA sample with 1ul dye in each wall, 120V for 40 minutes, 1 and 6 are ladder, the...
13 April 2023 6,351 3 View
Hello! I am struggling with Western blot. I would like to measure protein levels in bacteria cells, so I used western blot technique. Onto gel, I loaded bacteria lysates and the purified protein...
12 April 2023 2,147 6 View
When an ShRNA Knock down efficiency is not good, how I should improve this as I know ShRNA KD efficiency is not dose dependent.. it is meaningless to increase the amount of ShRNA right?
12 April 2023 9,814 4 View
Recently in one of our tests we identified a cellular protein expressed without any mRNA expression of the same protein intracellularly. What could be possible explanation? Kindly add possible...
11 April 2023 4,479 6 View
Hello! I am trying to use BTX ECM830 electroporator to introduce plasmid DNA into mammalian cells. I have tried using DMEM, HEPES-Based buffer and PBS, without success. i have used between 50 and...
10 April 2023 7,465 5 View
I drop-fixed fresh mouse brain tissue in a large volume (20:1 ratio) of 10% formalin for 16 hours before paraffin-embedding and slicing. Then I followed the standard IHC-codetection RNAscope...
08 April 2023 2,393 1 View
Hi Dear…………….Greetings………Those Are Not Arrogant Question And I Am Not Insulting You…..I Know Some Of Those Little Mystery…..…We Can Share Our Knowledge…...Do You Know Sky Mystery? Do You Know Who...
08 April 2023 4,055 3 View
I had done an amplification of pet28a plasmid to introduce some mutations in it using the designed primers. Pcr product was gel eluted, then sequentially was treated with Dpn1 and PNK enzymes....
08 April 2023 6,065 5 View
I have ready added RNase to the buffer, but still facing the problem. Can anyone please suggest me why? Picture has been attached (this 100bp band is more visible in some plasmids)
07 April 2023 8,165 2 View
I am currently working on Co-IP of NRIP and MyoD by overexpressing both proteins in HEK293T cell. The question is I cannot get a specific band of MyoD. For my input, I think the protein level is...
07 April 2023 2,505 4 View
Why my plasmid DNA gets degraded in Neb 10x buffer (2.1), used in restriction digestion? What can be the most probable reason for this?
06 April 2023 2,132 2 View
I am using Trizol reagent for RNA Extraction This is nanodrop evaluation after 1/10 dilution Is this curve accepted
05 April 2023 780 3 View
I want to overexpress PD-L1 in HEK293 to do immunoprecipitation. I designed a pcDNA3.1 within N-terminal 6xHis full-length PD-L1 and transfected it into HEK293T by PEI. I used RIPA to get 293...
04 April 2023 6,352 4 View
I've been having issues with my mouse genotyping results. I normally make a 1.5% Agarose Gel and add Ethidium Bromide to the solution to be able to image the results. I let all my samples run for...
04 April 2023 137 7 View
I tried human primers without any luck. Bovine primer worked for gapdh but did not work for dll4. I am repeating my experiment to confirm. Sheep primers are not easily available. any thoughts?
04 April 2023 3,452 3 View
Dear Professors, I am interested in using CANOCO software for my research but have difficulty accessing it. I would appreciate any advice on where I can download (for free) a copy of the...
03 April 2023 7,803 4 View
My cell lysates are so dilute that to load 20ug of protein into my gel, the sample volume is too big for the wells. I have heard of using MWCO filters, and I have had suggestions to let the cell...
03 April 2023 9,748 4 View
Can anyone tell me is there any way by which a single base can be changed in rice by using CRISPR Cas9
02 April 2023 5,197 0 View
Dear researchers, I am now trying to express several proteins from bacteria(Agrobacterium Tumefaceins) in one of its biosynthetic gene clusters. However, some of the proteins can be easily...
01 April 2023 569 4 View
Hello! When trying to transform E. coli DH10B or TOP10 with a large plasmid, like more than 65k bp, what do you think is the best time for the cells to be let recovering during the recovery time...
01 April 2023 8,884 2 View
I am using Phos-tag SDS PAGE and subequent western blotting to determine if the protein I am researching is being phosphorylated. I am using ECL to image my blots. However, I am getting really...
31 March 2023 4,950 3 View
Hi, I am using PBS, donkey serum and Tween for blocking solution. Unfortunately , the next day there is fungal contamination in the solution. How can I avoid this contamination for ICC
30 March 2023 6,849 1 View
Hi all, I've done WGS using nanopore, and have been able to successfully see 4 circularized plasmids and a full circularized chromosome. How do I know that these are the full plasmids without...
30 March 2023 8,959 2 View
I have been regularly amplifying bacteria with my plasmid of interest and performing midipreps yielding 400-600 ng/uL. As of late, my yields have been very low tanking to almost 80 ng/uL. I...
30 March 2023 6,396 3 View