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

Shortcomings or consequences of biological synthesis of silver nanoparticles?

Here I want to ask about the shortcomings and consequences of the biological synthesis of nanomaterials. Especially plant and microbial synthesis. Suggest me some reference paper. Thank you

20 December 2020 8,987 3 View

I would like to ask what if I perform an extra CaCl2 treatment to make competent cells?

I would like to ask that I have tried to make competent cells by 0.1MgCl2 and 0.1mM of CaCl2 treatments. My question is, what if I perform a real quick washing of bacterial pallet with 0.1mM CaCl2...

07 October 2020 7,274 6 View

Is there anybody in US have a Prep chiral Column to separate a racemic compound ?

I am looking for sombody in US who use a chiral prep column to separate a synthesis racemic H-tetrazol compound? I will send him the synthetic materials and the successful separation conditions...

16 August 2020 4,356 2 View

How to measure the frequency of transducer for ultrasonic welding of non-woven face mask?

Dear scientific community, We are trying to assimilate the non-woven ultrasonic welding machine for face masks. We are having some problems in generating enough heat to melt non-woven fabric. How...

07 May 2020 6,646 3 View

How can I make a .csv file that could be used in "Genetics" package implemented in R?

Dear all. Hello. I am trying to self learn the usage of r in genetical analysis. Can anyone tell me structure of .csv file which can be read in R from where I can start learning various executions...

10 April 2020 1,173 3 View

What should be the ideal graph of Co and Cross polarization for E and H-plane for a newly designed antenna?

What should be the ideal value(graph) of Cross and Co polarization for a designed antenna in HFSS? I am confuse about these two terms if anyone can explain it then please help me. Thanks.

09 January 2020 3,045 3 View

How to design a mix ratio of concrete for 40 MPa of compressive strength without using coarse aggregates?

I need a mix design which can give me 40MPa compressive strength concrete without using coarse aggregates. I am only using sand, water, fly ash, cement and superplasticizer for the mix design....

16 October 2019 3,562 7 View

What would you suggest me before running PCR reaction?

Dear Scientists, Hopefully, you all are well. My work is on pathogen related gene family in plants. I am performing PCR from last 4 months and getting results on and off. I am wondering that...

23 June 2019 7,166 8 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 View

RNA-seq DEGs downstream analysis and reporting in literature?

I used EdgeR to get the list of DEGs (differentially expressed genes) from my liver RNA-seq experiment. I have done the gene overrepresentation and GSEA based pathway analysis. I am currently...

23 February 2021 8,969 1 View

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

23 February 2021 7,326 5 View

RNA degraded after good nanodrop readings?

23 February 2021 8,850 2 View

Is there a way to extract total RNA and protein from cell culture at the same time, without denaturing protein?

Hi all, Is there a way to extract RNA and protein from the same cell culture sample? Because we need to use the protein for gelatin zymography at later time, we are looking for a method that can...

22 February 2021 5,341 5 View