Rezarf

... I used both 3M NaCl and 3M NaOAc in different samples and spun them down and removed the supernantant, the pellet in the NaOAc was much better than the NaCl which was more difficult to extract the supernatant without contamination. Also after washing with Isopropanol and 70% ethanol I managed to obtain a pellet of DNA, I added 100ul of TE buffer to resuspend and left it overnight in a shaker at 20 Celcius, but this morning only 1 of the 4 samples had resuspended.

Hi, Had a look at the isolation protocol, but I see it uses 1ml ethanol and 40ul Sodium acetate for Protein precipitation. I have been told the lab does not have phenol:chloroform or Ammonium acetate. Could i use Sodium Chloride or Sodium Acetate and would i still have to add in a protease? Thanks Rezarf

... was looking into phenol extraction but was asked to find an alternative reagent. Yes it is chromosomal DNA i wish to extract. Will have a look for those books. Been trying to find good protocols on-line but there are so many. I will also be extracting Chromosomal DNA from Human cheek and plant cells. I assume pulse-field electrophoresis is a bit more specialised than standard agarose gel, ie the equipment needed. Is there any over methods that could be used? Rezarf.

I am currently looking into the extraction of DNA from E. coli. I want to extract the DNA using a manual method and alsousing centrifuge. Once I have extracted the DNA I want to test it for yield,purity and possibly run gel electrophoresis to determine molecular weight. I have found some protocols online but I am stuck on a fewthings. Uses the centrifuge method a couple of this are not well explained. They use a Lysis solution which is SDS and also use RNasesolution. But the protocol also uses a Protein Precipitation Solution (PPS) anda DNA Rehydration Solution (RH) but does not say what it is made up of. Can anyone help clear this up for me? As once the DNA has been extracted I want to test for purityin the spectrophotometer. Thanks Rezarf

Hi, I am Rezarf and I am a HND student studying BioMedical Science. I am interested in DNA extraction just now and I am looking for ways of extracting DNA in the lab using E. coli, Human cheek cells and plant tissue. So anyone with any good ideas or information just let me know thanks. From sunny Scotland, where we get all 4 season in 1 day, lol. Rezarf