Anyone that can help with details in phosphopeptides enrichment?

Anyone know the amount of protein before enzymatic digestion for phosphopeptides enrichment of serume sample in Ti-IMAC? Has anyone ever use Ti-IMAC(the material is from Chinese company named...

28 February 2021 7,310 3 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

Is this the write procedure for acid digestion of soil for heavy metal analysis using AAS?

How to do acid digestion of soil sample with 3:1 aquaregia in an open vessel?

23 February 2021 3,053 7 View

Sanger Sequencing results not matching experimental results of Restriction Digestion, colony PCR and protein expression?

I have cloned two domains of a gene into two bacterial vectors using cut and paste cloning with different RE sites. I checked the putative clones by colony PCR and Restriction Digestion (fallouts...

18 February 2021 6,736 4 View

Alternatives to traditional cloning of insert into backbone (dsDNA)?

Hi, I am stuck with a cloning experiment: The insert & backbone(bb) share one compatible end at 3' end(NotI) and one incompatible end at 5' end (XbaI for insert; HindIII for backbone). So i...

16 February 2021 1,664 3 View

How to measure pH in anaerobic digester?

I'm planning on doing research by using anaerobic digester with liquid substrate and want to see its pH changes periodically. Any suggestion on measuring the pH with maintaining its anaerobic...

09 February 2021 1,293 1 View

Why RFLP Digestion looks in this way?

01 February 2021 9,576 3 View

Can I use 500ng of DNA for restriction digestion?

The recommended amount of DNA for restriction digestion is 1ug in a 50ul rx I've got around 500-600ng of DNA in a total volume of 35ul. Would I get satisfactory results with this amount of DNA for...

31 January 2021 729 6 View

After cutting bands having low molecular weight peptides obtained from SDS-PAGE,is it possible to elute these peptide(s) without digesting in trypsin?

Basically, I want to get the sequence of peptides. I assume, digesting these peptides from each band may give me a different sequence than the original through LC-MS analysis. If digestion doesn't...

30 January 2021 5,482 3 View

How to get the insoluble fraction of PK digestion for mitochondria?

I am trying to check for some proteins if they are located in the inner or outer mitochondria membrane after treating the cells with a neurotoxin. I am using PK to digest the mitochondria, but I...

27 January 2021 5,145 2 View