Could I use the TOP10 instead DH5alpha for pET-22b(+) plasmid transformation?

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Which is the optimal OD for protein induction?

My protein is expressed at 20C for 18h with 0.1mM IPTG Anyone can suggest optimal OD for induction?

10 November 2015 3,970 6 View

Can anyone suggest commercial vectors for secreted recombinant proteins to medium?

Which vectors are effective for secreted proteins from E. coli cell into medium?

10 November 2015 2,786 1 View

How do I express a secreted protein at 30-37’C?

What is the optimal IPTG concentration and incubation time after induction? Normally, My protein were induced with 0.5mM at 25'C for O/N but now I have limitation about shaker, so i'll to try new...

08 September 2015 1,141 4 View

Did anyone express rare codon in BL21DE3?

I wonder... What happen when express rare codon in E. coli? and Can be express full length protein even contain rare codon? ^-^

07 August 2015 3,985 0 View

Why my protein give a very light signal on SDS-PAGE?

My protein show correct size but very light on SDS-PAGE. In addition, Other size band is more bulky than my protein. I'm express in BL21(DE3) at 25'C for 18hr after added 0.5mM IPTG because...

06 July 2015 9,742 5 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

Low-yield gel extraction problem?

I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...

05 August 2024 9,798 3 View

The question is how to use Wavenet transform?

HOW CAN I WRITE A CODE TO USE THE WAVENET TRANSFORM AS A FEATURE EXTRACTION METHOD INSTEAD OF DWT IN MATLAB?

03 August 2024 7,829 0 View

Some new emerging problems on application of RL for scheduling in IoT networks?

I have seen plenty of existing works on applied Reinforcement Learning (RL) policies for optimized scheduling in IoT networks including Q-learning, DQNs, and the newer ones including PPO for...

01 August 2024 8,754 2 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

How to retain the a GFP tagged gene expression in stable cell line?

Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....

29 July 2024 5,007 0 View

What is the relationship between protein structure and N or C terminal tagging choosing?

I want to do 2,3-butanediol dehydrogenase(BDH) enzyme purification to confirm its activity for 2,3-butanediol. Before that, I need to confirm which N or C terminal tagging is better for enzyme...

28 July 2024 366 3 View

Why cannot i find my protein on cell surface after antibiotic selection of expressing plasmid?

I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...

28 July 2024 3,178 3 View

How does the Retention time effect the Biogas Production?

List down the various factors responsible and how does this effect the overall biogas production when it is related to Hydraulic retention time of the digester based on the feedstock?

27 July 2024 4,800 4 View

Can anyone help me with in providing the procedure for purification of GSTtagged protein by gst affinity chromatography without an automated machine?

Purification of gst tagged protein

25 July 2024 1,720 0 View