Which online software is reliable for Protein Transmembrane helix prediction

04 May 2017 6,200 5 View

Double band for a single protein on western blot?

Im working on yeast secretary protein, my protein was his tagged so i use Anti-his monoclonal antibody for WB, i can see the signal but i got two band on each protein (totally i have two proteins...

11 December 2015 5,327 12 View

What is the difference between Zymography and SDS-PAGE?

For determining the protein molecular weight, normally SDS-PAGE was used but some researchers also use zymography. im confused with both things. please clarify the difference between these two...

05 June 2014 8,429 6 View

Why should we use MgCl2 in particular instead of other chemicals for PCR?

Why should we use MgCl2 in particular instead of other chemicals for PCR?

02 March 2014 3,158 7 View

How to select a correct primer for an unknown organism for PCR?

We isolated bacteria from various samples. It is identified by biochemical tests as Lactobacillus spp. Now we are going to sequencing it for species level identification. I'm a little confused...

02 March 2014 6,089 6 View

Can anybody explain the target organism for pasteurization?

In milk numerous organisms grow well, that's why we pasteurize milk to remove unwanted microbes and their spores. But what is the exact target organism for pasteurization?

01 February 2014 1,002 10 View

How to find a specific gene responsible for particular charecter from the 16S rRNA sequence?

I need to identify the gene responsible for production of bacteriocin, and cloned it into another organism but I don't know how to identify the gene from 16S rRNA sequence.

01 February 2014 2,120 2 View

HPLC Peak has disappeared after one month. Any suggestions as to why?

We extracted secondary metabolite from wild Streptomyces spp. Initially, while running HPLC, peaks was obtained. However, after a few months, the peak disappeared (R.T 5.9-6.4 mins). Any suggestions?

31 December 2013 827 20 View

Is there any record of cholesterol reduction by Lactobacilus oris previously?

There are lots of studies which records the cholesterol reduction by Various Lactobacillus spp. but i want to know is there any researchers showing cholesterol reduction by Lactobacillus oris.

31 December 2013 2,836 1 View

Anybody explain about Metabolomics?

I want to know about the metabolomics, if which field it is used...

31 December 2013 6,660 0 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

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

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

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 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

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 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