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

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

website,genome sequence

01 February 2019 7,240 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 9,166 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 4,377 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 6,316 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 633 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 5,654 2 View

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

Tx6119( M15 containing fragment of SwpA26-226 from TX82 cloned into pQE30),when i run it as control without injecting IPTG i never saw any band on SDS-gel.

09 October 2017 3,269 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 5,763 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 401 4 View

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

Positive and negative control

05 June 2017 6,000 3 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

Why does my protein refolded to beta sheet during thermal denaturation analysis?

Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...

06 August 2024 1,989 3 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

How to use Density Functional Theory to calculate carrier mobilities of solid system?

Hello, everyone. I have tried to determine carrier motilities of some materials, by Density Functional Theory, using Quantum ESPRESSO. There are a few methods to do it, like a package called...

04 August 2024 8,894 1 View

Does long-term culture of murine Monocytes result in macrophage differentiation?

Hi, I am isolating monocytes from the bone marrow using the Mouse Monocyte EasySep kit. I want to treat these cells and monitor expression of specific markers over the course of 10 days. I will...

04 August 2024 7,282 2 View

How can I get my Granzyme B flow cytometry stain to be consistent?

I have used PE and PE-Dazzle 594 fluorochromes and have managed to get NK cells to properly show GranzymeB expression after 4 hr PMA/ionomycin stimulaton, but for some reason my CD8 cells in the...

01 August 2024 7,677 2 View

What is the best practice for resuspending cell pellets during competent cell preparation (both electro and comp)?

I'm fairly new to making homemade comp cells and have done so with some mild success. What is generally an acceptable way of resuspending comp cells during wash steps? I've been told to swirl the...

31 July 2024 8,309 0 View

Best mask design for metal deposition aiming build a Perovskite solar cell device?

What types of mask designs for metal deposition, to be used in a PVD system, are best suited for perovskite-based solar cells?

31 July 2024 4,835 3 View

How to culture and maintain Lx-2 cells?

Hi there! I recently started working on Lx-2 cells. When I thaw the cells, they start attaching within few hours but later in about 2 days they started rounding and dying. I used DMEM+2% FBS...

31 July 2024 3,893 0 View