I want to do protein expression in E. faecalis JH22 by using anhydrotetracycline (ATc) to induce protein expression .Need protocol for expression?

I want to use E. faecalis JH22 as competent cells to express my protein using anhydrotetracycline (ATc) to induce protein expression i need some standard protocol if someone has use it before.

06 July 2019 9,492 2 View

How to check the genome sequence of locus A of Enterococcus feacium TX16 species;any site for this ?

website,genome sequence

01 February 2019 7,163 2 View

When i purified my digested plasmid from the agrose gel ;and try to run of the gel again it just float from the well.Is my plasmid is contaminated ?

is my plasmid is contaminated with ethanol;how to avoid this contaminated

31 December 2017 4,285 0 View

Why are my gel electrophroesis samples are not moving?

I run gel electrophoresis to confirm digestion of my plasmid however my samples are remaining at the start of the gel. I have remade wash buffers and carrier buffers but am still not seeing my...

31 December 2017 6,241 0 View

POPINF plasmid i want to send for sequencing i donot what type of primer i have to use company have standard primers?

can anyone please recomend me any primer

10 November 2017 554 1 View

Ammonium sulphate precipitation method for protein , after centrifugation when i got the pellet,in which buffer dialysis of pellet has done to NMR?

in which i resuspend my pellet against which buffer i have to dialysis.

10 November 2017 5,575 2 View

I want to express my TX6119 strain ;i run it as control without injecting IPTG i saw no band?

09 October 2017 3,186 4 View

I am using Sybre Safe to stain the gel electropheresis;i 5microlitre in each welll but donot got any band, i just saw ladder band ,i am adding 6x dy?

Can anyone suggest why i am not getting any bands

09 October 2017 5,599 3 View

How much i add amipicillin and kanamycin into pQE30 (Qiagen) contains cloned fragment and transformed into M15 ( (pREP4) cells to do miniprep?

TX6119- Fragments were cloned into the expressionvector pQE30 (Qiagen) and transformed into M15 (pREP4) cells(Qiagen).i want to make culture in LB broth to do miniprep i am confuse how much i add...

07 August 2017 310 4 View

What type of positive and negative control;can be use for Histag antibody in Western blot?

05 June 2017 5,923 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

PUC19 as a transfer plasmid for lentiviral delivery?

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

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

How to get the vector of a specific vibration mode in phonon dispersion?

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

Specify the Burgers vectors for dislocations, which cannot be uniquely identified.?

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 am working on image transmission, i converted the image into binary vector, i want to extract the MSB of each pixel ?

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

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