Why plasmid after scale up not same of original?

I have cloned gene in one plasmid and sequence confirmed with Miniprep. I want to scale up same plasmid, for that I transformed my Final Miniprep plasmid into DH5 alpha cells and further used a...

11 December 2016 4,555 5 View

How to religate double digested plasmid?

Hi all, I have XbaI/BamHI digested linearized plasmid. I want to re ligate the same plasmid. Please provide suggestions.

06 July 2016 3,751 5 View

What are the important precautions to get minimum amount of endotoxin during plasmid isolation?

05 June 2016 9,557 4 View

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05 June 2016 8,212 5 View

What will be the qualitative and quantitative difference in plasmid scaled up from DH5 alpha (E.coli) and Stbl2 (E.coli)?

05 June 2016 936 5 View

Why the plasmid not linearizing with NruI restriction enzyme?

I am trying to linearize one circular plasmid with NEB NruI restriction enzyme. I have sequenced my plasmid and confirmed the NruI restriction site. I am using NEB buffer 3.1 in the NruI digestion...

05 June 2016 5,316 5 View

What is the highet transformation efficiency of DH5 alpha (E.coli )get to anyone ever?

If the number of colonies are very high (even hard to count on plate) when transformation done with 1 ng of DNA then how to calculate transformation efficiency? Anyone experienced that even with...

05 June 2016 5,007 4 View

What is the alternative for factor Xa to cleave the MBP tag from proteins expressed in pMAL system?

07 August 2015 4,831 4 View

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...

03 March 2021 5,978 5 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

Does Napabucasin soluble in dichloromethane and in ethanol?

I need to prepare a solution of Napabucasin (inhibitor of cancer cell stemness) in dichloromethane. The drug is expensive, so maybe somebody already tried to do it. If it is soluble, what is the...

02 March 2021 9,945 3 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

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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

Why am i not getting solid after reacting APTES with salicylaldehyde?

The reaction of APTES with salicylaldehyde in ethanol leads to the formation of imine compound yellow solid which is mention in literature, but I am not getting yellow solid I don't know why? when...

28 February 2021 2,955 2 View

How can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View