How to dissolve the precipitated DNA after EtOH purification? Should I increase the washes in the purification step?

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I was running Gaussian 09W Programmer in Frequency analysis but Chk File is not opened ?

gaussian frequency analysis was completed but chk file not open, Alpha MO energies Missing in view of the display.

24 January 2021 1,529 2 View

How to choose a perfect glass for solar still?

I need to select a transparent glass for solar still. but i know the specification all. but i need high transmissivity glass. but i dont know what brand , and what thickness i should choose. i...

13 November 2019 9,799 4 View

How much is 1 volume of 70% ethanol?

I need to isolate RNA from the tissue sample. The procedure is given like "add 1 volume of 70% ethanol to the lysate, and mix well by pipetting". How much ethanol should I add?

19 November 2017 8,272 5 View

RAW cell culture and FACS?

Hi folks, I am growing RAW 264.7 cells in 12 well plates, RPMI with FBS and PS. When harvested and acquired cells in FACS, in few of the stimulated wells population is missing or second population...

23 August 2016 9,977 3 View

How can we predict material swelling in single point diamond turning?

In SPDT material swelling increases the surface roughness of the work piece. How can we predict that the material going to swell for given machining parameters?

06 August 2012 4,354 2 View

How are bioinformatics and computational biology tools used for genetic inputs to study hematopoietic lineages?

How to understand vaarious genetic inputs that control the development of various hematopoietic lineages employing various molecular, genetic, bioinformatic and computational approaches.

18 July 2012 2,780 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

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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 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

Is there a method of determining feed digestibility without the need to re-pellet the feed?

I am aiming to determine the ADC values of a commercial fish feed, but I do not have a steam pelleter or extruder available to me. Ideally, the pellets could be coated with some inert marker....

28 February 2021 7,967 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

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