Why E. coli BL21 strain can not be used for plasmid preps?

Why E. coli BL21 strain can not be used for plasmid preps?

01 February 2015 9,576 0 View

Is there any chemical difference in Heparin resin used in HiTrap Heparin HP and Heparin sepharose 6 FF from GE healthcare?

Hitrap Heparin HP is coupled to the Sepharose High Performance base matrix via the N-hydroxysuccinamide coupling method and in case of Heparin sepharose 6 FF, Heparin is linked to the Sepharose...

31 December 2014 6,439 1 View

Can we sequence DNA on sequencing gels using fluorescent labels?

Can fluorescent labels be detected with a phosphor imager? We have Fujifilm- FLA 9000 and Biorad Screen Eraser-K set up for detection.

10 November 2014 8,774 3 View

Can we use M13 Reverse and M13 Forward (generally used for sequencing insert in pUC18 MCS) for seuencing/amplifying fragment of M13mp18?

Do these two primers have a sequence in M13mp18 DNA, as the name suggests that these are from M13? Please provide an explanation.

06 July 2014 6,939 1 View

How can I remove DNA from bacterial crude lysates?

06 July 2014 698 12 View

Does anybody know of a plasmid or DNA vector which lacks or has a single TTAA site?

As it is a 4 bp sequence, it frequently occurs in most plasmids. If anybody knows of such plasmid lacking or having single TTAA site?

02 March 2014 7,422 1 View

How do we equilibrate the millipore protein concentrator while using it for the first time?

Can it be reused, and if yes, how many times?

01 February 2014 5,205 8 View

Can coomassie stained gels which have been destained, be stained again? With coomassie stain and silver stain?

I have stained my SDS-PAGE with coomassie blue but bands are very light. Probably, the stain was not good. Can the gel be stained again with new coomassie or silver stain? Will the staining be...

01 February 2014 8,373 12 View

Do proteins retain activity after acetone precipitation?

I need to concentrate my protein samples for running SDS-PAGE as well as checking activity.

01 February 2014 9,457 5 View

What is the binding capacity of DEAE cellulose?

Concentration of sample is 10-12 mg/ml and I am planning to use 4 ml DEAE cellulose resin. I need to know the amount of protein sample required to be loaded on the column to get good resolution?

01 February 2014 4,914 5 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 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

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 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