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 purify digested PCR product or Plasmid in more concentartion?

I am trying to purify my Digested PCR product and Plasmid with Promega Kit.The purified product is of low conc.

11 December 2018 2,903 0 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

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 to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 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 simultaneously perform multiple biochemical assays?

I am a research scholar working on heavy metal stress on plants. I will be doing biochemical characterisation( protein, carbohydrate, proline, antioxidant enzymes and many more assays) at interval...

01 March 2021 6,999 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

Bought HUVEC from Invitrgen.need to revive it.expected it is cryofreezed in DMSO.Do I need to remove the DMSO for plating the cell for the 1st time?

After I cryofreeze the cell do I need to remove the DMSO for culturing the cell for next passages .

28 February 2021 7,738 3 View

POlymer+DMSO solution and moisture??

What could be the reason of moisture at the sides of Vial when I am preparing my polmyer+DMSO solution for membrane fabrication? What etiquettes I am possibly missing here?

28 February 2021 6,516 5 View

I have a time series data of some biochemical studies ( DNA and Chromosomal damage) which I intend to use to further predict into future ?

I have a time series data of some biochemical studies ( DNA and Chromosomal damage) which I intend to use to further predict into future without necessarily conducting the assay for an extended...

23 February 2021 7,842 1 View

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

Why anti-CD20 or other antibodies are not used to label target cells in flow cytometry t-cell cytotoxicity assay?

New to the field of cell therapy. Why fluorescent-labeled antibodies to target cell antigens are not used for determining target cell count by flow cytometry in cytotoxicity assay (eg car-t)? Cell...

21 February 2021 3,278 3 View