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

I want to use PET-Duet vector but i donot protocol how to do restriction cutting with two enzymes ;if i want to insert two gene of interest?

As PET duet vector is for cloning of two gene of interest;i want the protocol for it.

06 July 2018 9,061 0 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

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

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

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Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 View

How to perform statistical analysis of coefficients extracted from transform techniques?

I am studying the potential of DCT and FFT coefficients extracted from a signal to classify the different objects that emit the signal. I applied the Shapiro-Wilk to confirm that data was not...

01 March 2021 8,079 2 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

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 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 improve RNA isolation?

Hi, I have been struggling with a weird issue for my RNA samples. I used the Qiagen RNA plus mini kit. For some reason no matter how careful I am with the washes, when I measure the RNA, I get...

22 February 2021 1,286 6 View

I have a problem with kanamycin - why the kanamycin doesn't affect?

Hi everyone, I need to expression of GFP with pET28 plasmid in E. coli BL21(DE3) plysE. Transformed cells have kanamycin and chloramphenicol resistance. Normally, only transformed cells should...

22 February 2021 4,032 4 View