A. C. E. Draper

5 Questions 3 Answers 0 Followers

Questions related from A. C. E. Draper

Do lentiviruses containing DNA simply integrate this in to host cells?

This is probably a daft question; I have cloned in a gene of interest to a lentiviral expression vector and successfully made lentiviral particles that easily transduce HEK cells producing the...

03 December 2018 3,747 1 View

Does anyone have a good protocol for fractionation of nuclear from cytoplasmic proteins?

My current protocol is: Procedure: Note: All centrifugations should be done at 4°C. 1) Scrape cells on 10 cm plates using 5 ml of cold PBS plates immediately and place in 15 ml falcon tube. 2)...

29 November 2016 5,515 3 View

Does a low TU/ml in a specific lentivirus equate to a low transduction efficiency?

There is quite a range in the TU/ml between different batches of virus that I make. If I make 4 different gene variants at a time using the same plasmid backbone, packaging and envelope plasmids...

13 June 2016 8,470 9 View

Does variation in lentiviral pellet colour after ultracentrifugation matter?

I am making 2nd generation lentiviruses by transfecting HEK cells with the following; pMD2V, pPAX-2 and a lentiviral plasmid vector. I harvest the media at 48 and 72 hours, and then spin (4500g...

16 January 2015 7,688 0 View

Is the use of the Clontech Lenti-X 4th generation lentiviral system essential for lentiviral production using pLVX-AcGFP-C1 lentivectors?

Having produced a fusion protein lentiviral vector (GFP in-frame with the gene of interest), using the Clontech pLVX-AcGFP-C1 vector I now want to produce these in to a lentiviral system. However...

03 November 2014 6,113 2 View