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

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

Protein-aptamer gel electrophoresis help, please??

Hi, I have problems with running gel electrophoresis. I have tried agarose gel electrophoresis and native PAGE. I have two proteins, which have molecular weights of ~30kDa and ~180kDa and two...

03 March 2021 4,275 4 View

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

How to regulate SSCP temperature in simple PAGE?

I'd like to perform single-strand conformation polymorphism (SSCP) in my thesis, however I cannot control the temperature of the vertical PAGE since we are using the conventional tanks. Is there a...

02 March 2021 9,157 1 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

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 2 View

Why are my bands weird in this Agarose gel?

Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.

01 March 2021 9,952 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

What do I do with DLS peaks that are from the buffer?

I'm looking at the aggregation of my protein sample using DLS. Unfortunately, my buffer (20mM HEPES) also results in a set of peaks. These are at approximately 1 and 1000 d.nm. The lowest peak...

01 March 2021 9,015 2 View