Hiya *Half an hour in the library saves a month of research.* That's what my late advisor Dr. Frolow used to say a lot and he was not wrong.
Erenpreisa J. 1981. Staining of DNA with uranlyacetate in hydrolyzed ultrathin sections. Acta histochem 68(1): 22-26 there are several very clever methods, using depurination and various chemical treatments (e.g. phenylhydrazine) followed by staining with various heavy atoms (tungsten derivatives) In general, shadowing with tungsten is a highly adequate method, see for example gorgeous images of replicaiton forks here: https://www.jbc.org/content/285/18/13349.full.pdf Artem - Cosmic Cats approve of this message On Tue, Aug 4, 2020 at 7:11 AM Panne, Daniel (Prof.) < daniel.pa...@leicester.ac.uk> wrote: > Hi Meytal, > > 1. The concentration seems way too high at 254mg/ml. Typically 0.1-5mg/ml > are used. > 2. Uranyl ions react with phosphate groups resulting in positive stain. > The resulting contrast is frequently poor. > 3. Rotary shadowing can be used to enhance contrast. > > Best, > Daniel > > > On 4 Aug 2020, at 11:20, Marin van Heel < > 0000057a89ab08a1-dmarc-requ...@jiscmail.ac.uk> wrote: > > > Dear Meytal, > > Suggestion: Apply a strong high-pass filter to the images before visually > judging them! > > Marin > > On Tue, Aug 4, 2020 at 3:50 AM Meytal Galilee <meytal.gali...@gmail.com> > wrote: > >> Hi Arunabh Athreya, >> Sure (this is the linear 2200 b.p. at 175uM) >> <image.png> >> >> >> >> Get Outlook for Android >> <https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Faka.ms%2Fghei36&data=02%7C01%7Cdaniel.panne%40leicester.ac.uk%7C1d84ebc5b8674ad1d62b08d838601d22%7Caebecd6a31d44b0195ce8274afe853d9%7C0%7C1%7C637321332736273265&sdata=1JpwGUyNpGCO3IlNY8y0eXKdGulww020ZzJLc93idfo%3D&reserved=0> >> >> >> ------------------------------ >> *From:* Arunabh Athreya <arun...@iisc.ac.in> >> *Sent:* Tuesday, August 4, 2020, 12:54 >> *To:* CCP4BB@JISCMAIL.AC.UK; Meytal Galilee >> *Subject:* Re: Advice on DNA negative staining >> >> Hi Meytal >> >> Do you have an EM image snapped from your negative staining sample? >> >> Get Outlook for Android >> <https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Faka.ms%2Fghei36&data=02%7C01%7Cdaniel.panne%40leicester.ac.uk%7C1d84ebc5b8674ad1d62b08d838601d22%7Caebecd6a31d44b0195ce8274afe853d9%7C0%7C1%7C637321332736273265&sdata=1JpwGUyNpGCO3IlNY8y0eXKdGulww020ZzJLc93idfo%3D&reserved=0> >> >> >> >> ------------------------------ >> >> To unsubscribe from the CCP4BB list, click the following link: >> https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 >> <https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2FWA-JISC.exe%3FSUBED1%3DCCP4BB%26A%3D1&data=02%7C01%7Cdaniel.panne%40leicester.ac.uk%7C1d84ebc5b8674ad1d62b08d838601d22%7Caebecd6a31d44b0195ce8274afe853d9%7C0%7C1%7C637321332736283221&sdata=QooLJZLuHXjOPYtWFVQXnLB4CShQRxG3u5EX%2FACb%2FuQ%3D&reserved=0> >> > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > <https://eur03.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2FWA-JISC.exe%3FSUBED1%3DCCP4BB%26A%3D1&data=02%7C01%7Cdaniel.panne%40leicester.ac.uk%7C1d84ebc5b8674ad1d62b08d838601d22%7Caebecd6a31d44b0195ce8274afe853d9%7C0%7C1%7C637321332736283221&sdata=QooLJZLuHXjOPYtWFVQXnLB4CShQRxG3u5EX%2FACb%2FuQ%3D&reserved=0> > > > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
