Re: [ccp4bb] AW: [ccp4bb] crystals that dont diffract :( :(
Dear Careina, Please make sure that your crystals are protein or salt, possibly using Izit dye.Note: there is no correlation so far hypothesized on the diffraction of ugly / beautiful / needle / plate type crystalsI believe! With best regardsS.M.Jaimohan PhD On Tuesday, 14 August, 2018, 7:21:09 PM IST, ferrer wrote: On 14/08/2018 12:27, Careina Edgooms wrote: Cryoprotectant was 50% PEG just added to the buffer that it crystallised in. It crystallised under batch. which kind of batch plate ? Some are not suitable to in situ. JL On Tuesday, August 14, 2018, 12:11:17 PM GMT+2, Hughes, Jon wrote: maybe it's the cryobuffer that's the problem (you didn't mention it). you could try to fish the crystals with minimal liquid attached by mounting them in oil rather than a cryobuffer. or you could test the native diffraction "in situ" (at room temperature in the drop): quite a few beamlines offer this possibility these days. best jon Von: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] Im Auftrag von Careina Edgooms Gesendet: Dienstag, 14. August 2018 11:59 An: CCP4BB@JISCMAIL.AC.UK Betreff: [ccp4bb] crystals that dont diffract :( :( I got the most beautiful crystals I have ever seen and they don't diffract at all. Not poor diffraction, NO diffraction. Anyone know why this could be and how I can go about fixing it? I had three beautiful crystals and not one diffracted. I did leave them in the drop for about 3 weeks before harvesting and in liquid nitrogen for about a month before diffracting. Could that be a factor? If I regrew more beautiful crystals and diffracted straight away could that help? Careina To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 -- Jean-Luc Ferrer Institut de Biologie Structurale 71 Avenue des Martyrs CS 10090 38044 Grenoble Cedex 9 - FRANCE Ph.: +33 (0)4 57 42 85 22 Cell: +33 (0)6 89 45 13 57 email: jean-luc.fer...@ibs.fr To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
Re: [ccp4bb] AW: [ccp4bb] crystals that dont diffract :( :(
On 14/08/2018 12:27, Careina Edgooms wrote: Cryoprotectant was 50% PEG just added to the buffer that it crystallised in. It crystallised under batch. which kind of batch plate ? Some are not suitable to in situ. JL On Tuesday, August 14, 2018, 12:11:17 PM GMT+2, Hughes, Jon wrote: maybe it's the cryobuffer that's the problem (you didn't mention it). you could try to fish the crystals with minimal liquid attached by mounting them in oil rather than a cryobuffer. or you could test the native diffraction "in situ" (at room temperature in the drop): quite a few beamlines offer this possibility these days. best jon *Von:*CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] *Im Auftrag von *Careina Edgooms *Gesendet:* Dienstag, 14. August 2018 11:59 *An:* CCP4BB@JISCMAIL.AC.UK *Betreff:* [ccp4bb] crystals that dont diffract :( :( I got the most beautiful crystals I have ever seen and they don't diffract at all. Not poor diffraction, NO diffraction. Anyone know why this could be and how I can go about fixing it? I had three beautiful crystals and not one diffracted. I did leave them in the drop for about 3 weeks before harvesting and in liquid nitrogen for about a month before diffracting. Could that be a factor? If I regrew more beautiful crystals and diffracted straight away could that help? Careina To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 -- Jean-Luc Ferrer Institut de Biologie Structurale 71 Avenue des Martyrs CS 10090 38044 Grenoble Cedex 9 - FRANCE Ph.: +33 (0)4 57 42 85 22 Cell: +33 (0)6 89 45 13 57 email: jean-luc.fer...@ibs.fr To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
Re: [ccp4bb] AW: [ccp4bb] crystals that dont diffract :( :(
Dear Careina,
Unfortunately there is no direct connection between nice looking crystals and
good diffraction.
Sometimes ugly crystals can diffract well while nice ones don’t.
You may try crystal dehydration ; it allows to remove excess of solvent and
change crystal packing which can lead to better crystal order and better
diffraction.
Here are some examples described in the literature
https://www.ncbi.nlm.nih.gov/pubmed/24311593
https://www.ncbi.nlm.nih.gov/pubmed/22232185
https://www.ncbi.nlm.nih.gov/pubmed/12575933
Bests,
Zaineb
Le mardi 14 août 2018 à 12:11:23 UTC+2, Hughes, Jon
a écrit :
#yiv5746868635 #yiv5746868635 -- _filtered #yiv5746868635
{font-family:Helvetica;panose-1:2 11 6 4 2 2 2 2 2 4;} _filtered #yiv5746868635
{panose-1:2 4 5 3 5 4 6 3 2 4;} _filtered #yiv5746868635
{font-family:Calibri;panose-1:2 15 5 2 2 2 4 3 2 4;}#yiv5746868635
#yiv5746868635 p.yiv5746868635MsoNormal, #yiv5746868635
li.yiv5746868635MsoNormal, #yiv5746868635 div.yiv5746868635MsoNormal
{margin:0cm;margin-bottom:.0001pt;font-size:12.0pt;font-family:New
serif;}#yiv5746868635 a:link, #yiv5746868635 span.yiv5746868635MsoHyperlink
{color:blue;text-decoration:underline;}#yiv5746868635 a:visited, #yiv5746868635
span.yiv5746868635MsoHyperlinkFollowed
{color:purple;text-decoration:underline;}#yiv5746868635 p
{margin-right:0cm;margin-left:0cm;font-size:12.0pt;font-family:New
serif;}#yiv5746868635 p.yiv5746868635msonormal0, #yiv5746868635
li.yiv5746868635msonormal0, #yiv5746868635 div.yiv5746868635msonormal0
{margin-right:0cm;margin-left:0cm;font-size:12.0pt;font-family:New
serif;}#yiv5746868635 span.yiv5746868635E-MailFormatvorlage19
{font-family:sans-serif;color:#2E74B5;}#yiv5746868635
.yiv5746868635MsoChpDefault {font-size:10.0pt;} _filtered #yiv5746868635
{margin:70.85pt 70.85pt 2.0cm 70.85pt;}#yiv5746868635
div.yiv5746868635WordSection1 {}#yiv5746868635
maybe it's the cryobuffer that's the problem (you didn't mention it). you could
try to fish the crystals with minimal liquid attached by mounting them in oil
rather than a cryobuffer. or you could test the native diffraction "in situ"
(at room temperature in the drop): quite a few beamlines offer this possibility
these days.
best
jon
Von: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK]Im Auftrag von Careina
Edgooms
Gesendet: Dienstag, 14. August 2018 11:59
An: CCP4BB@JISCMAIL.AC.UK
Betreff: [ccp4bb] crystals that dont diffract :( :(
I got the most beautiful crystals I have ever seen and they don't diffract at
all. Not poor diffraction, NO diffraction. Anyone know why this could be and
how I can go about fixing it? I had three beautiful crystals and not one
diffracted. I did leave them in the drop for about 3 weeks before harvesting
and in liquid nitrogen for about a month before diffracting. Could that be a
factor? If I regrew more beautiful crystals and diffracted straight away could
that help?
Careina
To unsubscribe from the CCP4BB list, click the following link:
https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
To unsubscribe from the CCP4BB list, click the following link:
https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
To unsubscribe from the CCP4BB list, click the following link:
https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
Re: [ccp4bb] AW: [ccp4bb] crystals that dont diffract :( :(
Cryoprotectant was 50% PEG just added to the buffer that it crystallised in.
It crystallised under batch.
On Tuesday, August 14, 2018, 12:11:17 PM GMT+2, Hughes, Jon
wrote:
#yiv2688348338 #yiv2688348338 -- _filtered #yiv2688348338
{font-family:Helvetica;panose-1:2 11 6 4 2 2 2 2 2 4;} _filtered #yiv2688348338
{panose-1:2 4 5 3 5 4 6 3 2 4;} _filtered #yiv2688348338
{font-family:Calibri;panose-1:2 15 5 2 2 2 4 3 2 4;}#yiv2688348338
#yiv2688348338 p.yiv2688348338MsoNormal, #yiv2688348338
li.yiv2688348338MsoNormal, #yiv2688348338 div.yiv2688348338MsoNormal
{margin:0cm;margin-bottom:.0001pt;font-size:12.0pt;font-family:New
serif;}#yiv2688348338 a:link, #yiv2688348338 span.yiv2688348338MsoHyperlink
{color:blue;text-decoration:underline;}#yiv2688348338 a:visited, #yiv2688348338
span.yiv2688348338MsoHyperlinkFollowed
{color:purple;text-decoration:underline;}#yiv2688348338 p
{margin-right:0cm;margin-left:0cm;font-size:12.0pt;font-family:New
serif;}#yiv2688348338 p.yiv2688348338msonormal0, #yiv2688348338
li.yiv2688348338msonormal0, #yiv2688348338 div.yiv2688348338msonormal0
{margin-right:0cm;margin-left:0cm;font-size:12.0pt;font-family:New
serif;}#yiv2688348338 span.yiv2688348338E-MailFormatvorlage19
{font-family:sans-serif;color:#2E74B5;}#yiv2688348338
.yiv2688348338MsoChpDefault {font-size:10.0pt;} _filtered #yiv2688348338
{margin:70.85pt 70.85pt 2.0cm 70.85pt;}#yiv2688348338
div.yiv2688348338WordSection1 {}#yiv2688348338
maybe it's the cryobuffer that's the problem (you didn't mention it). you could
try to fish the crystals with minimal liquid attached by mounting them in oil
rather than a cryobuffer. or you could test the native diffraction "in situ"
(at room temperature in the drop): quite a few beamlines offer this possibility
these days.
best
jon
Von: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK]Im Auftrag von Careina
Edgooms
Gesendet: Dienstag, 14. August 2018 11:59
An: CCP4BB@JISCMAIL.AC.UK
Betreff: [ccp4bb] crystals that dont diffract :( :(
I got the most beautiful crystals I have ever seen and they don't diffract at
all. Not poor diffraction, NO diffraction. Anyone know why this could be and
how I can go about fixing it? I had three beautiful crystals and not one
diffracted. I did leave them in the drop for about 3 weeks before harvesting
and in liquid nitrogen for about a month before diffracting. Could that be a
factor? If I regrew more beautiful crystals and diffracted straight away could
that help?
Careina
To unsubscribe from the CCP4BB list, click the following link:
https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
To unsubscribe from the CCP4BB list, click the following link:
https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
To unsubscribe from the CCP4BB list, click the following link:
https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
[ccp4bb] AW: [ccp4bb] crystals that dont diffract :( :(
maybe it's the cryobuffer that's the problem (you didn't mention it). you could try to fish the crystals with minimal liquid attached by mounting them in oil rather than a cryobuffer. or you could test the native diffraction "in situ" (at room temperature in the drop): quite a few beamlines offer this possibility these days. best jon Von: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] Im Auftrag von Careina Edgooms Gesendet: Dienstag, 14. August 2018 11:59 An: CCP4BB@JISCMAIL.AC.UK Betreff: [ccp4bb] crystals that dont diffract :( :( I got the most beautiful crystals I have ever seen and they don't diffract at all. Not poor diffraction, NO diffraction. Anyone know why this could be and how I can go about fixing it? I had three beautiful crystals and not one diffracted. I did leave them in the drop for about 3 weeks before harvesting and in liquid nitrogen for about a month before diffracting. Could that be a factor? If I regrew more beautiful crystals and diffracted straight away could that help? Careina To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
