Publishing in F1000Research??

Hi, Anyone has first hand experience of publishing in F1000Research. I guess it follows open peer review and does not encourage measurable parameters like impact factor! What are pros and cons...

05 June 2019 9,547 1 View

Protein-protein interaction??

Hi, I have two proteins that are his tagged at N-terminus. One of them forms hexamer around 370kDa and other is 17kDa. I want to check their interaction and get Kd value. Which of the following...

03 April 2019 1,474 5 View

Designing mutation experiments??

Hi all, I have a protein with conserved Threonine and one Lysine in the middle of the protein sequence. Being a charged and conserved amino acid across family domain of this protein, I have strong...

02 March 2019 4,847 5 View

E.Coli expression strain for expressing mycobacterial membrane protein gene?

Hi, I have cloned some three different constructs which are putative membrane protein of Mycobacterium tuberculosis into pET28a with N-term His tag. I tried to express them in BL21, Rosetta, C41,...

11 December 2018 1,871 3 View

Bioinformatic tool to predict flexible region in a protein?

Hi, Can anyone please tell some bioinformatics server that helps in predicting the location of flexible region in a protein.?

10 November 2018 6,510 0 View

Difference between Coomassie Brilliant Blue R-250 and G-250??

Hi, Can anyone please highlight the difference between Coomassie Brilliant Blue R-250 and G-250?? Which one of them will stain the SDS gel faster?

09 October 2018 2,780 0 View

Citing preprint articles?

Hi, I was just curious to know whether the scientific articles and review published in preprint repository like bioRxiv are citable? If yes, is it good practice to do that since mostly such...

09 October 2018 4,686 4 View

How to degass a buffer with detergent?

Hi, I want to add about 5 CMC DDM detergent in my Size exclusion buffer. Is it advisable to add detergent and then degass by vacuum or degass the buffer first and then add DDM dropwise slowly??

08 September 2018 4,062 3 View

1M NaOH cleaning overnight?

Hi, One of my colleague put the Superdex 200 10/300 GL column mistakenly in 1M NaOH at flowrate of 0.2ml/min overnight instead of water wash. Will the column be stable? How to know that the...

07 August 2018 5,171 1 View

Publishing in bioRxiv??

Hi, Can someone please tell pros and cons briefly of publishing in preprint servers like bioRxiv?

07 August 2018 7,602 1 View

Waters 2707 HPLC autosampler problem?

I am using a 2707 waters HPLC device. When I try to inject a sample, it says missing plate or rack. I changed the needle and calibrated its position but I still get the same problem. I even get...

02 March 2021 1,408 1 View

Should we change media after1st 6hrs after plating HUVEC bought from INVITROGEN?it's not recomended by invitrogen.Will DMSO present in stock matter?

We have HUVEC bought from Invitrogen. Cat no -C-003-5c. while first plating the cells we have bought the protocol suggests not to centrifuge the cell with media to remove the DMSO. Should we...

02 March 2021 3,713 2 View

How to normalise data on OPM package from R Studio ?

Good afternoon, I recently used OmniLog from BIOLOG for my experimentations : I tested the metabolism of different strains on 2 types of plates. I have 16 strains of 3 different groups...

02 March 2021 3,584 1 View

How to grow E.coli on the low salt LB agar plate with 50 or 100ug/ml of Blasticidin?

Hello, I would like to ask how to grow E.coli which resists 50 or 100ug/ml of Blasticidin. I have tried many methods by using low salt Luria-Bertain(LB) agar plate. However, no colonies grow. Let...

01 March 2021 2,027 2 View

Benefits of the 24 welled plate?

We have always used the 96 welled plate, the 48 welled plate but apparently there is much evidence on the 24 welled plate and above all its benefits.It states it is the same you would use on ELISA...

28 February 2021 1,149 1 View

Can bacterial suspension be pipetted directly onto petri dish and agar poured onto it?

Hi, recently I encountered a protocol for plating which was not the usual method I have been practicing (pour LB agar onto plate, allow it to solidify, pipette bacterial suspension onto plate,...

28 February 2021 7,292 3 View

What is the best surface to culture cells isolated from a skin biopsy?

Hello, recently I am having some difficulties with maintaining the cell culture of primary cells isolated from a skin biopsy (normal and malignant). The coating of culture plates with Matrigel (a...

26 February 2021 6,151 2 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Can cells be reused after using WST 8/ CCK 8 from Sigma Aldrich?

Hi all, Can cells be reused after I have seed them onto 96 well plates and test them with cell counting kit at OD450 (CCK8) at 12 hours, wash them with PBS x3 and add new media and let them...

24 February 2021 5,072 1 View

Standard curve shifting over time. How do I stabilise the ELISA?

24 February 2021 1,365 3 View