Kinetic and thermodynamics study of cloned alpha amylase

05 June 2013 5,786 2 View

Protein purification

I have expressed my protein in E. coli. and I want to purify it. I performed the ammonium sulfate precipitation with 15%, 25%....75% in the indicial samples and ran both the supernatant and...

03 April 2013 6,049 6 View

Universal primers for 16s PCR ribotyping

We have isolated a bacterium from dung for the biogas production. Biochemical tests indicated that it could be Methanosarcina. I want to perform 16s PCR-Ribotyping. Can any one suggest the...

02 March 2012 628 3 View

Can linear DNA be used as template for site-directed mutagenesis?

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

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Can anyone provide the Support vector regression code structure of LIBSVM MATLAB or python ?

Please, if Gaussian Parameter optimization is possible in this code structure

28 February 2021 278 3 View

Does anyone have a sequence for the human CD34 promoter?

I'm looking for the human CD34 promoter sequence to engineer into a vector construct. Can anyone recommend a good source for this sequence? Thanks very much, Laura

25 February 2021 8,205 1 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

Why are Sanger Sequencing reads interrupted in some edited clones?

I have a Crispr/Cas9 edited bulk population and I can briefly check the indel ratios in the bulk population using Sanger sequencing. After cloning single cells, I wanted to check the indel types...

18 February 2021 8,198 6 View

Hi, can anybody help me with the suggestions how can I amplify a stress induced gene of 1107 bp?

I am trying to amplify the TGA1 (Transcription factor 1 ) of Arabidopsis ,which is 1107 bp. I have treated the plants with Salycylic solution over night and extracted the RNA and make cDNA. I...

17 February 2021 7,802 2 View

Alternatives to traditional cloning of insert into backbone (dsDNA)?

Hi, I am stuck with a cloning experiment: The insert & backbone(bb) share one compatible end at 3' end(NotI) and one incompatible end at 5' end (XbaI for insert; HindIII for backbone). So i...

16 February 2021 1,664 3 View