What is the standard protocol in doing replicates for C. elegans lifespan assay on solid media (thru worm picking)?

I would like to know what is the standard protocol in the number of worms per plate and number of plates per concentration of extract X in a lifespan assay done in solid media and not automated...

23 February 2021 9,933 1 View

Native gel blot unable to to observe antibody binding?

Hello, I'm trying to utilize a native gel for a protein dimer experiment. Everything seemed to work relatively well until I try to examine the blot in a ChemiDoc MP. I don't see any antibody...

30 September 2020 6,059 3 View

Isolates that grow on CHROMagar col-APSE will not grow on MacConkey agar supplemented with Colistin Sulphate?

I prepare the CHROMagar Col-APSE (for selection of colistin resistant gram-negative bacteria) exactly to manufacturers guidelines. I have E. coli isolates that grow perfectly well on this media,...

26 March 2020 9,377 4 View

DNA gel has a U shape?

22 August 2017 4,812 3 View

Can anyone recommend a proper procedure using mitomyocin C to inactivate mice T-cells in order to perform a proper mixed lymphocyte reaction?

This MLR will be done using two different strains of mice that is why it is necessary to inactivate the T-cells, so that possible proliferation can be observed.

06 October 2016 2,257 3 View

Can anyone help me with my a stent and balloon inflation model in Abaqus?

22 November 2015 6,911 15 View

Can transient transfection using COS-7 cells with a plasmid encoding a neomycin resistance be used a positive transfected cell selection method?

I want to do a transient transfection using COS-7 cells with a plasmid encoding a neomycin resistance, can I use this as a positive transfected cell selection method? COS-7 cells transfect with...

04 March 2015 9,231 5 View

Are cos-7 cells good for patch clamp?

I have been trying to patch transfected cos-7 cells (Kv channel plasmid) and am having an incredibly difficult time getting a seal let alone breaking in. Has anyone every patched with these cells...

10 December 2014 3,117 2 View

How do I optimize PCR reactants?

Hi everyone! Attached you will a EtBR gel showing failed PCR results in lanes 2 and 3, with positive results for a DNA ladder in lane 1 (100 bases). Additionally, I have attached a photo of the...

01 January 1970 3,528 3 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

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

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

I have a problem with kanamycin - why the kanamycin doesn't affect?

Hi everyone, I need to expression of GFP with pET28 plasmid in E. coli BL21(DE3) plysE. Transformed cells have kanamycin and chloramphenicol resistance. Normally, only transformed cells should...

22 February 2021 4,032 4 View

How to successfully revive cells stored at -80 degree C for a year?

I am working on C6 cells along with other neuronal cell lines. We have preserved the cells in 90%FBS and 10% DMSO solution. We are reviving the cells after a year and cells are either not binding...

18 February 2021 4,499 1 View

What are alternatives for B27+ Supplement in neuronal cell culture?

Hello everybody In our iPSC-derived neuronal culture we use Maturationmedium, supplemented with B27+. As Thermo Fisher run out of stock and can deliver earliest in May, I wanted to ask whether...

16 February 2021 3,348 1 View

Can 5-FOA Diffuse Through Agar?

I am preparing to do a gene knockout in the filamentous fungi Coccidioides posadasii targeting pyrG/URA3 so that 5-FOA can be used for selection pressure instead of traditional antibiotic...

16 February 2021 169 4 View

How do I change g/l into mM for bacto agar. I am trying to get the millimolar content of sodium and chloride percentage in bacto agar. How do I do it?

I am doing a salt tolerance-related experiment and for that I need to relate my salt levels to that in bacto agar. I don't want my salt concentrations to increase due to already salt content...

09 February 2021 4,158 1 View

RWP1 growing in RPMI medium?

Hello everyone, Has anyone try to culture the normal prostate RWPE-1 cell line in RPMI medium supplemented with foetal bovine serum? There's some articles available but neither describe details...

08 February 2021 6,455 1 View

Transformation into the Stbl3 chemo competent cells?

03 February 2021 1,111 5 View