Why my Gibson assembly doesn't work?

I'm assembling 3 PCR fragments with my vector pBAD and transforming them into dh5a. The second PCR fragment has 4 variations to it and the template for this PCR fragment comes from a plasmid. I've...

01 February 2019 5,696 3 View

Does anyone have a good protocol for flagella preparation/purification?

I need a protocol to shear flagella off E.coli O157:H7. I have been using a protocol online that is basically resuspending an overnight culture in a syringe and then, spinning it down to acquire...

02 March 2017 6,637 2 View

How does one crystallise a protein with the presence of TCEP as the reducing agent, and with it's co-factors (NAD+ and Fe2+)?

I have a protein which I know is stable with the presence of co-factors (NAD+ and Fe2+). Recently i have been trying to purify it with the TCEP as a reducing agent since my protein is known to...

10 November 2016 185 2 View

Why mbp-cleaved protein disappears after anionic exchange?

I am purifying a protein (47 kDa) that is fused onto an MBP (42 kDa). The PIs for both proteins are 6.48 and 4.9 respectively. After purifying it using a dextrose column, I have buckets of fused...

04 May 2016 8,945 8 View

How to find self-associating motifs in a protein sequence?

04 May 2016 3,195 3 View

Why did the cleaving of my MBP tag on my protein failed?

I'm purifying a protein that is fused onto an MBP tag (42.5 kDa). My protein size is roughly the similar size of the tag (47kDa). Together, they form a complex about 89 kDa. After a run through a...

03 April 2016 8,667 3 View

How do I know if a flagella is in R or L conformation?

I have sequenced my sample and found some mutaions in the fliC gene. I have found that they have introduced some aa changes however, I would like to know if this changes the conformation into R or...

02 March 2016 7,471 1 View

How to generate GNOM file (.out) for DAMMIF runs?

I have .dat files from scatter that contain the Dmax of a protein and now I want to generate ab initio models. however I dont know how to generate .out files that are needed byy DAMMIF programs on...

09 October 2015 9,090 1 View

Why does the pressure alarm keeps going off during Nickel column purification?

I'm using a new column (5 ml Ni NTA HiFliQ column from Generon) to purify my protein AdhE which is Histagged and sized 96 kDa. However, during purification, the alarm pressure keeps going off even...

07 August 2015 5,183 10 View

Why are there no protein expressed even under different expressing conditions?

I am purifying AdhE (96 kDa) protein which is bifunctional and ontains 2 functional domains; (acetaldehyde dehydrogenase and Alcohol dehydrogenase). Before purification, I did a small scale...

07 August 2015 4,143 7 View

What is the most Operating System (OS) which can be INSTALL on Tiny hard devices like Raspberry Pi (RPi) hardware ?

I'm targeting to deploy a mesh network and manually configure MANET routing protocols. I'm preparing scenarios, architectures, and hard devices needed to do that. Are there some step-by-step...

03 March 2021 1,931 5 View

Which of the Covid-19 Vaccines would elicit the best immunoprotection and why?

I want to know which of the available Covid-19 Vaccines would elicit the best immunological response and why

02 March 2021 1,388 3 View

How can I integrate a routing protocol in a tiny device (e.g. Raspberry Pi)?

Hi Community, I'm facing the issue of integration/compiling a new routing protocol in a WSN simulator. The final goal is to successfully add, configure this routing protocol in hardware devices...

01 March 2021 9,332 6 View

Experimental Design: How deal with pseudreplicates with immunological and clinical-chemical paramaeters?

Hello everybody, I am investigating the influence of a toxinon various clinical-chemical and haematological parameters in fish. I have three treatments (control, low dose, high dose) in my...

24 February 2021 1,243 1 View

Looking for advices and tips to improve the staining and the gating strategy in human blood?

Hi, I am staining a human blood with CD19, CD3,CD14 and CD16. My gating strategy in FlowJo goes like this: - First, I gate a general population SSC-A VS FSC-A => Then gate for single cells, =>...

21 February 2021 5,724 4 View

How can I add a new WSN routing protocol in NS2 or NS3?

I'm trying to find in literature a way to help me adding a modified Yen's algorithm in ns2 or its variance ns3 or another simulator. Have you some tips which can be helpful, I'm opening in the...

19 February 2021 7,981 3 View

I want to simulate a privacy-preserving algorithm in MQTT for IoT devices, but I'm not sure which simulation software is best suited for this?

I want to apply a privacy-preserving algorithm in MQTT for IoT devices. Which tool do you suggest to simulate this algorithm? I want to measure the performance on the IoT battery device? Actually,...

28 January 2021 9,561 3 View

How can we simulate vehicle to infrastructure communication in NS-3?

I am working on handoff management in IEEE 802.11p/WAVE based VANETs. I want to know if I can simulate V2I communication mode using NS-3, which will lead to the simulation of handoff process. Most...

16 January 2021 1,733 4 View

Can you polarize Jurkat cells into different T helper subsets?

I am wondering if anyone has had success with in vitro polarization (via cytokines) of Jurkat cells. My goal is to understand the affect of a drug on Treg induction and differentiation. I...

12 January 2021 3,606 2 View

Is there anyone who can suggest some related work based on HMM implementation for routing protocol in WSNs?

I'm trying to solve the energy issue conservation in WSNs and I want to use the Markov technique to predict this conservation and demand the routing protocol to select an optimal path among a...

05 January 2021 1,994 2 View