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Questions related to Vectorization
Hi, I am trying a cloning over a long time. My vector is of about 4.7 kb and insert 1.4 kb. I am using Xho1 and Kpn1 as restrictions. I am attaching some files for details. In the first image I...
05 September 2021 3,610 5 View
I am trying to clone/insert a second gene to the pRSFDuet1 vector but the second gene is not showing up in the gel after cloning. What precautions should be taken to successfully clone the second...
05 September 2021 8,290 1 View
Hello. I recently cloned some plasmids for shRNA. I ordered PAGE-purified shRNA duplex oligo, and inserted it in lentivirus vector for shRNA(addgene cat.no. 8403, pLKO.1-puro) using AgeI and MluI...
03 September 2021 4,978 10 View
What is the appropriate wavelength for measuring Streptomyces spp growth
03 September 2021 9,135 2 View
Hello dear friends I am working on gene cloning. One of the stages is pCAMBIA 1304 vector extraction. The low concentration of vector at the final stage of pCAMBIA 1304 vector extraction, is my...
31 August 2021 2,873 3 View
I am trying to digest pET28a by Bam HI and Xho I. I am using 1.5 ug DNA and 1ul of enzymes (which ia 20 unit per ul) and incubated at 37*c for 6 hours and in second reaction, incubated for 2...
29 August 2021 6,225 5 View
Dear All, I have implemented an1D DOA based on a MUSIC algorithm. How can I go from 1D to 2D DOA, my particular problem is implementing 2D steering vector and how to perform the search through...
28 August 2021 3,840 2 View
Considering a matrix A which has vectors v1=[1;0;0] and v2=[1;1;0] i.e. this matrix A is spanned by the vectors v1 and v2. The rank of this matrix A is 2. Going by the definition of the rank of...
25 August 2021 8,481 8 View
I carried out digestion of vector pLentiv2 (14kb) with restriction enzyme BsmBI at 37C for 3 hours, and performed a gel extraction to purify the cut DNA fragment for cloning. I ran both the uncut...
22 August 2021 3,475 1 View
In the case of decomposition based MaOPs, for example NSGA3, some predefined Ref.Points are associated to the population members in order to decompose a Many Objective problems to the series of...
21 August 2021 3,863 4 View
I intend to do CRISPR KO. Can I insert gRNA into lentiCRISPR V2 via infusion cloning? Also, will the whole sequence of lentiCRISPR V2 be incorporated into host genome or only partially? Where...
21 August 2021 7,748 2 View
Several papers reported that the mode effective refractive index (neff) of vortex modes (such as OAM+1 mode) would be changed with the variation of different pitches (or twist rates) in the...
18 August 2021 3,143 3 View
I am cloning Newcastle disease virus fusion gene into vector PC DNA-3.1 but problem occurs as in last part after digestion when i run plasmid DNA ON GEL only band of vector or some time no band...
15 August 2021 959 2 View
Hi, This is a brief description of my latest head scratcher: I am digesting a vector with XhoI and HindIII-HF (both from NEB), then I dephosphorylate the digested vector with the Antarctic...
14 August 2021 4,355 3 View
Can we utilize the N-terminal His-tag of pET32b vector for His tag purification? I have incorporated the stop codon in my reverse primer. Thank you.
11 August 2021 4,387 2 View
Have anyone tried to clone a gene into gateway destination vector via restriction enzyme cloning? I know LR cloning is good but the LR cloning kit is kind of expensive to our lab. So I was...
08 August 2021 1,038 4 View
Dear all, I am now facing a problem. After ligation of a 12000 bp vector with 2000 bp gene fragment, no clones can be obtained after performing DH5alpha transformation? Therefore, are there any...
04 August 2021 1,135 4 View
I was constructing marker free vector for Pichia pastoris , but I got a problem when adding CRE gene under AOX promotor which is showing leaky expression. Please help me with ur valuable suggestions ?
03 August 2021 2,985 2 View
Hello, everyone. I used a transgenic vector for SCNT and successfully obtained cloned individuals. I screened the cloned individuals for the presence of vector by PCR and distributed individuals...
03 August 2021 9,482 4 View
Can anyone please guide me through how to design crRNA for Cas12a based detectioin. For IVT purpose where should be the T7 promoter and where I should add repeats.
02 August 2021 5,165 1 View
Hello researchers ! so I have a gene with a GFP fluorophore that was cloned inside of it with BamH1 restriction sites. I wanted to replace GFP with mCherry but the thing is this enzyme site is not...
20 July 2021 3,972 14 View
According to the description on TAIR, the vector inserted in WiscDsLoxHs mutants (Arabidopsis) is pDsLoxHs. However, I cannot find the sequence of this vector, I am asking for help. Thanks.
14 July 2021 9,123 2 View
Thanks in advance.
06 July 2021 896 9 View
Poynting vectors in curl free vector fields were discussed in a previous thread as possible sources of propulsion including several types. When Poynting vectors are also canceled out, the result...
06 July 2021 6,706 13 View