I would like to know if is possible to re amplify and PCR fragment for Gibson Assembly ?
I did cut and purified the gel band but the amount in ng/uL is not enough for Gibson Assembly later.
Anthropometrically and conceptually, association does not equate causation. However, regarding anthropometric indicators (i.e., metrics of general and abdominal obesity), most of studies worldwide...
20 April 2024 8,779 6 View
I need to evaluate viral accumulation by qPCR in plants with DNA and RNA virus coinfections . I dont know how proceed in tge sample treatment before the qPCR . Please someone has done this before ?
04 December 2023 3,651 0 View
I have been culturing human microvascular endothelial cells (HMEC-1) on my own-made low-serum media. The cells expand fine in wells and flasks T25, T75, and T180 using 0.2 ml/cm2, so the media is...
23 November 2023 1,439 1 View
Im running a Gibson Assembly, using two fragments of 2750 pb and a vector of 3300pb. Then when I transform in DH5alfa or 10 beta got a higher band than I expected. Also I did PCR from miniprep and...
03 December 2022 3,356 0 View
I have flow cytometry data of T cells for which I measured the expression of 5 exhaustion markers. I would like to know how many of my cells simultaneously express any combination of 1, 2, 3, 4 or...
17 July 2022 4,584 2 View
I would like to use the SKBR3 cell line in a long term co-culture with engineered T cells. To reduce their proliferative capacity and prevent rapid depletion of the media I would like to irradiate...
06 April 2022 5,533 0 View
I'm designing primers for Gibson Assembly . My main query about that is the harpain and self dimer formation ? Did you have problems with that ?
20 January 2022 2,727 5 View
Dear all, I have recently come across several examples in Social Sciences where supervisors become co-authors of published articles by MSc and PhD students. While I understand a supervisor might...
07 October 2018 2,915 94 View
I am using Chloroquine and its effect on autophagy and I want to treat my cell culture with a dose close to the exposition with CQ treatment. So how valid is to use the mean serum concentration?...
23 November 2017 2,245 2 View
How to establish the appropriate reactions and innteractions between compounds developed in the produce's senescence and colorimetric compounds for the sensor of an intelligent packaging...
14 October 2017 8,587 3 View
We assume this to be true. We also assume that the vacuum bomb is the latest version of explosives with an explosive power of a few to ten kg of TNT equivalent. It has the unique characteristic of...
04 August 2024 4,534 1 View
Given that the bacterial genome has over 800 contigs, but its quality metrics are good, with a completeness of 98.55% and a contamination of 0.68% as assessed by CheckM, what specific validation...
01 August 2024 1,514 1 View
We are in need of primers for a PCR protocol for chicken genome. We are looking at the COI barcode, 648-bp fragment of the 5' end of COI. Specifically Primers FalcFA, BirdR1, CO1_ExtF,...
16 July 2024 118 2 View
I'm currently working on gel purification in order to get DNA fragment for T4 ligation. But I have been able to take only 8.5ng/μl as a highest concentration of DNA fragment. And could not get...
01 July 2024 4,912 9 View
Please, can the fragmentation obtained from the mass spectrometry be predicted more easily using any free software? Kindly recommend it, thanks.
01 July 2024 5,327 6 View
Hello, I am trying to clone 1334 bp insert into 6.1 kb vector using HiFi DNA assmebly/Gibson assembly kit. I am taking 100ng of vector and 1:5 molar ratio of vector: insert and 4 hrs of...
27 June 2024 9,652 2 View
To do gibson assembly cloning can I use DH5alpha strain of E. coli. for transformation?
20 June 2024 774 1 View
I am trying to do gibson assembly cloning. I have two fragments for gibson assembly cloning (plasmid backbone and insert). After gibson assembly reaction I have transformed this in E. coli....
19 June 2024 6,404 4 View
I have to join tungsten wire to metallic plate with that adhesive and entire assembly has to be heated upto 600C in ultra high vacuum. I know about zirconia paste but it is not readily available...
18 June 2024 3,092 3 View
Dear cloning community, Is it possible to use Overhangs with large Tm difference with NEB Builder HiFi DNA assembly? 5´ 26 bp Tm 57ºC 3´ 24 bp Tm 72ºC Unfortunately, extending the 5´ doesn't...
17 June 2024 8,552 2 View