My plasmid DNA is 96ng. How do i calculate the volume of bsa I (20000 U) to use to digest it?

More Benjamin Kurya's questions See All

What is the biochemistry of acidic solvents for phenolics extraction in strawberry?

I am using the Folin-Ciocalteu method for analysis of total phenolic content in ripe strawberries. I've read several protocol that suggest a methanol/H2O/formic acid (80:19:1) extraction solution....

22 February 2021 6,560 3 View

Are there any open-source datasets containing physiological information about exercises to be used for an OR-model?

I have just recently started a new "weekend project" in addition to my master's studies and I am looking for a data-set. I would like to use some Operations Research to design an optimal gym...

22 February 2021 8,023 1 View

How reliable is estimation of copy numbers by absolute qPCR?

Hello, I would like to compare the expression levels of several genes in insects, so I prepared standard curves and estimated copy numbers of each sequence in my samples. But for one of the genes,...

11 February 2021 8,421 13 View

Antibody fragments after Dha (Dehydroascorbic acid) re-oxidation?

Hi, I am trying to re-oxidize the disulfide bridges of an antibody with DHA. I always have some good amount of HL fragments (heavy + light chain fragment), despite varying reaction times, pH,...

31 January 2021 9,495 2 View

Please, any real time simulation tool recommendation for islanding detection and control in DG to me aside MATLAB as it keeps freezing and very slow?

I am currently working on islanding detection and control in distributed generations. I would kindly ask that you recommend a real-time simulation tool that helps me in this study. I have tried...

13 January 2021 3,626 3 View

Can decolonial theory speak to human rights movements?

Decolonial theory tends to avoid the language of human rights due to a concern about who is included in "the human" of human rights. But many struggles today start from the language of human...

20 December 2020 7,896 3 View

Ethanol or Isopropanol for insect preservation?

Hello, I am wondering whether Isopropanol (cheaper) can be used in the same way as ethanol for preservation and storage for molecular and DNA analyses, particularly for insect samples. Any ideas...

29 July 2020 4,994 17 View

Is it okay to have R-Squared or an Adjusted R-Squared to be over 90% in multiple linear regression? Does this raise any suspicion or question?

I ran multiple linear regression by using six IVs to predict household water insecurity in rural Ghana. My output came out with R-squared and Adjusted R-Squared of .981 and .962, respectively....

10 July 2020 5,854 2 View

What is the best program to simulate temperature of the tool tip during turning (Abaqus or Ansys)?

Dear all, I would like to simulate the temperature of a tooltip during turning. What program should I use best? And does anybody know where I can find a template where I can start from? This...

28 June 2020 2,572 3 View

No title

Hi i would like to know if there are some stat methods to aggregate Results of only one intervention group between different studies of a systematic review. in fact, I want to resume a binary...

12 June 2020 5,184 2 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 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

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

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

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

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Michael J. Benedik

If you have 96 ng of plasmid then in theory you need 0.1U of enzyme to digest in in an hour (assuming the unit is defined as 1 hour). Practically speaking if you add 1-2U you should be fine.

Katie A S Burnette

The product insert that comes with the enzyme will have a standard protocol for how much to use.

Do you mean you have exactly 96 nanogram of plasmid OR do you have a plasmid at the concentration of 96 nanograms per microliter?