I am using pPICZalfa vector with gene length 1.1 Kb. Currently I am trying for In-vitro multicopy generation (Bgl II - BamHI), but the transformation efficiency in DH5alfa is decreasing as my construct size is increasing. Should I try PTVA method?
Transfection efficiency in general decreases with increasing plasmid size.
Exactly! The plasmid size is increasing at every ligation step.
I have tried various methods using imageJ to measure mean fluorescence intensity of dapi stained nuclei but none of them seem to be working. Ive attached an image i am trying to measure the...
15 February 2021 8,637 3 View
I am using coupled transient thermal electric step to simulate thermal and electric flux load on cfrp layered composite. DFLUX been used for surface heat flux and URDFIL for element deletion but...
25 December 2020 2,551 1 View
Can anyone please explain this? How do I account for plastic hinges in my model and how do I modify the stiffness after plastic hinge is formed? Need a small example. Thank You
15 November 2020 1,967 3 View
I want to model rc element subjected to cyclic loading, How do I enter hysteresis data in the material model? I want to use Mander's 1988 model to get data for concrete. I dont know how to enter...
06 September 2020 7,651 3 View
I am working on a water quality model for the Mula Mutha river. I have a doubt regarding the dispersion coefficient to be entered. whether to enter a fixed value or computed value? what are...
22 May 2020 848 1 View
Why the final docked complex file output from ZDOCK is not compatible for Chimera?
24 April 2020 3,347 3 View
I tried finding frequency by various tools such as audacity (got frequency 0-16000 Hz). Tried with Librosa(Python package) and got the frequency of human recorded audio files in the range of...
06 April 2020 7,607 6 View
I am looking for software that will take input as a research paper (pdf format) and generate the data from plots and graph in it. as similar to NLP softwares.
04 February 2020 4,244 3 View
I have been using miRNA Isolation mini kit (qiagen 217004) for isolating RNA from PBMCs stored in RNA later at -80 C. The cell no. was 2 million cells. The RNA yields obtained are around 30-40...
15 October 2019 4,731 1 View
I did not not find the answer anywhere. Please help.
19 August 2019 9,950 3 View
Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...
03 March 2021 401 4 View
Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...
28 February 2021 4,868 2 View
For example, I have this phonon dispersion(Fig. 1), and now I would like get vector of atoms which describes TA1 in P. I know v_sim can show vibrational animation of specific mode, but I do not...
20 February 2021 4,332 4 View
1. A bcc metal single crystal was sliced along (1 -1 0) plane for TEM study. Assuming that the TEM available is equipped with a goniometer stage which tilts up to 45o. The Burgers vector of...
20 February 2021 2,370 1 View
i have a vector 1*512 size, i want to take the first value every 8 elements and put these elements at the end of the vector,how to do this on matlab ?
19 February 2021 2,352 5 View
Hello, I am planning to insert GOI between CaMV 35 promoter and mGFP in pCambia1302 to produce a fusion protein and see subcellular localization of the protein. But the vector says it's membrane...
15 February 2021 4,353 3 View
Hi Fellow Scientists! I am trying to clone my 300 bp insert into a 6 kb vector. I run a few different ligation reactions using (1:3 vector: insert molar ratios) 10, 20,30, and 40 ng/ul vector...
15 February 2021 9,563 6 View
Are E.coli strains M15 and pET28a vector systems compatible? Can I over-express my protein which is of plant origin. Previously I was using BL21 DE3 strains. Was confused to use M15 strains for...
10 February 2021 2,564 3 View
When co-transfect S2 cell with selection vector (pCoHygro or pCoBlast ) to make a stable cell line, does the gene integrate into the genome or just like E.coli? How do the plasmids amplify within...
09 February 2021 5,423 1 View
I have tried to clone a specific gene into pCAMBIA1304 vector. But all the attempts failed. I used ligation kit from both Promega Corporation and Takara Bio. However, there was one time I used...
04 February 2021 525 1 View