I purchased pdCas9 from addgene and want to clone and express it in pET22b vector. Please, I need a detailed explanation.
Also, where can I get purified dCas9 for purchase? Thanks.
O.J
PCR by designing primers of the dCas9 and target vector RE sites
or
Cut the dCas9 region from previous vector, repair the ends and ligate to new vector. But you have to care about in-frame codon.
https://www.researchgate.net/post/Is_it_possible_to_transfer_your_construct_of_interest_from_a_plasmid_to_another_one
following
As Golzar Hossain answered, you can do PCR or subcloning into a new vector. For subcloning you need to check the vector maps of the addgene vector and the new vector you want to design in order to use the proper restriction enzymes.
what can be the undiscovered benefits of applying telematic technologies for road pavement maintenance, incident reduction and environmental protection?
29 June 2024 7,507 1 View
most of the time when i search fo certain topic i found the human verification respond and nothing change there after
19 March 2024 6,870 0 View
Hello! I have a methodological question and I hope someone can help me. I collected data via an online survey with a sample size of 2600 (N = 2600). Initially, I aimed to examine the effect of...
26 February 2024 6,911 4 View
Hi RG Community I had input all the appropriate files, however, received an Action halted on error! My ORCA generated .gbw QM Output file was not recognized (please see screenshot attached). I...
15 February 2024 2,553 1 View
Typically, it is said that biofilter by their nature allow the settling of bacteria beneficial to the treatment of fishwaste. Fish culture though is known for producing high amounts of ammonia in...
28 January 2024 2,398 3 View
From what I can tell by searching the "Your World in Data" website, eastern Europe seems to have an expected lower than average all-cause mortality rate than the western first world nations, who...
04 January 2024 7,066 0 View
Hello RG Community:), I have a ligand exhibiting penalties between 10-50 within a .str file generated via a CGenFF, the generated .prm file does not exactly match the ligand atoms nor the closest...
25 December 2023 4,364 1 View
Hi I had modelled a heptamer transmembrane protein missing 10-residues within the extracellular loops via MODELLER constraining the known residues as a heptamer (when MODELLER modeled as a monomer...
08 December 2023 4,512 0 View
Hello towards the above topic, I successfully loop-modeled a transmembrane heptamer protein (426-residues per monomer) exhibiting 7-extracellular loops each missing 11-residues via MODELLER via...
03 December 2023 9,528 2 View
Need a simulation tool to generate synthetic data Specialized engine simulation software (for control and diagnosis) how and what to do 🙏?
16 November 2023 4,128 1 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...
05 August 2024 9,798 3 View
Dear Researchers I need to know, how to load and plot 2D-PIV (particle image velocimetry) recorded velocity vector field data in Tecplot? Thank You
02 August 2024 3,615 1 View
Hi, everyone. I want to transfer two genes in to the pseudomonas bacteria that isolated from the soil. The bacteria that I want to use for cloning haven't been identified and not be sequenced,...
02 August 2024 3,987 1 View
Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...
31 July 2024 9,892 4 View
I want to do 2,3-butanediol dehydrogenase(BDH) enzyme purification to confirm its activity for 2,3-butanediol. Before that, I need to confirm which N or C terminal tagging is better for enzyme...
28 July 2024 366 3 View
List down the various factors responsible and how does this effect the overall biogas production when it is related to Hydraulic retention time of the digester based on the feedstock?
27 July 2024 4,800 4 View
Purification of gst tagged protein
25 July 2024 1,720 0 View
Hello, We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't...
25 July 2024 4,927 3 View
The literature on Supply Chain Integration (SCI), Supply Chain Management (SCM), and collaborations, spanning several decades, deserves special attention. This topic, although addressed for a long...
23 July 2024 5,672 3 View