Protocol for extraction and derivitization of fatty acid for GC analyis?

I want to extract my P450 reaction mixture and derivitized for GC analysis. I have read about BSTFA method, but i have no idea about the procedure. Please help me to understand the procedure.

31 December 2018 6,860 12 View

How to fuse two independent domain?

I have two independent domain cloned in E.coli and i want to fuse them both just to get a single domain by PCR. How to get this by PCR?

09 October 2018 7,125 5 View

How to prepare 2% DMSO soln (v/v)?

I have to conduct assay for enzyme.For this, I have to dissolve my substrate(palmitic acid) 200mM in DMSO having conc 2% (v/v). Please help me .

09 October 2018 2,372 0 View

Facing problem in Cloning?

facing problem in cloning.No colony in transformants plate.

07 August 2018 9,241 10 View

Facing problem in substrate binding with BM3 protein?

During performing assay with BM3 protein (CYP 450),I am facing problem with binding of substrate.I am using Napthalene and Palmitic acid as a substrate.

06 July 2018 1,978 3 View

Facing problem in cloning using Bamh1 and Not1 as restriction sites?

I am using two enzymes BamH1 and Not1 to cut my insert and vector both.Aftr transformation,I am not getting any getting? Do Bamh1 not1 are not compatible ?

05 June 2018 4,541 1 View

Do we need to add potassium phosphate later in tb media?

Do we need to add potassium phosphate later in teraffic broth media ?

05 June 2017 7,302 0 View

Composition of sonication buffer for bacterial cell lysis ?

05 June 2017 6,251 8 View

Do we express our recombinant protein into TOP10 instead of BL21(DE)?

Do we express our recombinant protein into TOP10 instead of BL21(DE)? Is this affects its protein expression?

03 April 2017 7,309 3 View

I m trying to break bacillus megaterium cell with high pressure homogeniser..bt notng happen to my cell...i continued upto 2.30 hr?

Buffer composition 50mM tris hcl Nacl 150mM Mgcl2 5mM 10%glycerol

01 January 1970 3,802 3 View

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

Research on Supply Chain Management in Indian context?

Hi, I am planning to apply for the PhD degree in the Supply Chain Mgt. with specific area of "Cold Storage warehouses" during Pandemics and wars. Where lock downs and shut downs are frequent....

02 March 2021 285 2 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

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Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 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