[ccp4bb] DNA oligonucleotide purification
Title: DNA oligonucleotide purification All, Our laboratory is looking for a new column to purify short (40mers or less) DNA oligonucleotides that will be used for crystallization setups. I wanted to ask if anyone has suggestions of a column that will give excellent separation of N and N-1 products. We have a Shimadzu HPLC system. We are considering Dionex columns as well as Oligonucleotide Separation C18 columns from Waters, but if there are others that are better we would like to know. Any suggestions will be greatly appreciated. Thank you! Jamie Wallen
Re: [ccp4bb] DNA oligonucleotide purification
Hi Jamie - We've had good success with these columns from Dionex (the DNAPac PA-100 and the DNAPac PA200 (Analytical 4x250mm and Semi-Prep 9x250mm,)). We use them precisely for the application you describe. http://www.dionex.com/en-us/products/columns/bio/nucleic-acid/dnapac-pa100/l p-73370.html http://www.dionex.com/en-us/products/columns/bio/nucleic-acid/dnapac-pa200/l p-73371.html cheers, Kushol Kushol Gupta, Ph.D. Research Associate Van Duyne Laboratory - HHMI/Univ. of Pennsylvania School of Medicine BLOCKED::mailto:kgu...@stwing.upenn.edu kgu...@mail.med.upenn.edu 215-573-7260 / 267-259-0082 From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Wallen, Jamie Sent: Thursday, April 28, 2011 9:32 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] DNA oligonucleotide purification All, Our laboratory is looking for a new column to purify short (40mers or less) DNA oligonucleotides that will be used for crystallization setups. I wanted to ask if anyone has suggestions of a column that will give excellent separation of N and N-1 products. We have a Shimadzu HPLC system. We are considering Dionex columns as well as Oligonucleotide Separation C18 columns from Waters, but if there are others that are better we would like to know. Any suggestions will be greatly appreciated. Thank you! Jamie Wallen
Re: [ccp4bb] DNA oligonucleotide purification
Title: Re: [ccp4bb] DNA oligonucleotide purification Kushol, Thank you for the reply. Might I ask what kind of HPLC system you have? Our HPLC is old and has stainless steel pumps, and to our knowledge this is not compatible with the Dionex columns (due to metal leaching). Dionex has recommended either a titanium or peek system. As our HPLC is still running well, we were hoping to find a column that can tolerate the stainless steel. Jamie On 4/28/11 9:11 AM, Kushol Gupta kushol.gu...@gmail.com wrote: Hi Jamie Weve had good success with these columns from Dionex (the DNAPac PA-100 and the DNAPac PA200 (Analytical 4x250mm and Semi-Prep 9x250mm,)). We use them precisely for the application you describe. http://www.dionex.com/en-us/products/columns/bio/nucleic-acid/dnapac-pa100/lp-73370.html http://www.dionex.com/en-us/products/columns/bio/nucleic-acid/dnapac-pa200/lp-73371.html cheers, Kushol Kushol Gupta, Ph.D. Research Associate Van Duyne Laboratory - HHMI/Univ. of Pennsylvania School of Medicine kgu...@mail.med.upenn.edu BLOCKED::mailto:kgu...@stwing.upenn.edu 215-573-7260 / 267-259-0082 From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Wallen, Jamie Sent: Thursday, April 28, 2011 9:32 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] DNA oligonucleotide purification All, Our laboratory is looking for a new column to purify short (40mers or less) DNA oligonucleotides that will be used for crystallization setups. I wanted to ask if anyone has suggestions of a column that will give excellent separation of N and N-1 products. We have a Shimadzu HPLC system. We are considering Dionex columns as well as Oligonucleotide Separation C18 columns from Waters, but if there are others that are better we would like to know. Any suggestions will be greatly appreciated. Thank you! Jamie Wallen
Re: [ccp4bb] DNA oligonucleotide purification
We're using an older Dynamax system that is outfitted with PEEK throughout. Cheers, Kushol From: Wallen, Jamie [mailto:jwal...@biochem.wustl.edu] Sent: Thursday, April 28, 2011 10:17 AM To: Kushol Gupta; CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] DNA oligonucleotide purification Kushol, Thank you for the reply. Might I ask what kind of HPLC system you have? Our HPLC is old and has stainless steel pumps, and to our knowledge this is not compatible with the Dionex columns (due to metal leaching). Dionex has recommended either a titanium or peek system. As our HPLC is still running well, we were hoping to find a column that can tolerate the stainless steel. Jamie On 4/28/11 9:11 AM, Kushol Gupta kushol.gu...@gmail.com wrote: Hi Jamie - We've had good success with these columns from Dionex (the DNAPac PA-100 and the DNAPac PA200 (Analytical 4x250mm and Semi-Prep 9x250mm,)). We use them precisely for the application you describe. http://www.dionex.com/en-us/products/columns/bio/nucleic-acid/dnapac-pa100/l p-73370.html http://www.dionex.com/en-us/products/columns/bio/nucleic-acid/dnapac-pa200/l p-73371.html cheers, Kushol Kushol Gupta, Ph.D. Research Associate Van Duyne Laboratory - HHMI/Univ. of Pennsylvania School of Medicine kgu...@mail.med.upenn.edu BLOCKED::mailto:kgu...@stwing.upenn.edu 215-573-7260 / 267-259-0082 From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Wallen, Jamie Sent: Thursday, April 28, 2011 9:32 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] DNA oligonucleotide purification All, Our laboratory is looking for a new column to purify short (40mers or less) DNA oligonucleotides that will be used for crystallization setups. I wanted to ask if anyone has suggestions of a column that will give excellent separation of N and N-1 products. We have a Shimadzu HPLC system. We are considering Dionex columns as well as Oligonucleotide Separation C18 columns from Waters, but if there are others that are better we would like to know. Any suggestions will be greatly appreciated. Thank you! Jamie Wallen
Re: [ccp4bb] DNA oligonucleotide purification
We actually screen with unpurified oligos (up to just over 30nt) and it works just fine. Saves lots of time $$. If we get hopefull crystals, we pay for purification or do it ourselves by gel. Sometimes it makes the crystals better and sometimes it doesn't. Phoebe = Phoebe A. Rice Dept. of Biochemistry Molecular Biology The University of Chicago phone 773 834 1723 http://bmb.bsd.uchicago.edu/Faculty_and_Research/01_Faculty/01_Faculty_Alphabetically.php?faculty_id=123 http://www.rsc.org/shop/books/2008/9780854042722.asp Original message Date: Thu, 28 Apr 2011 08:32:23 -0500 From: CCP4 bulletin board CCP4BB@JISCMAIL.AC.UK (on behalf of Wallen, Jamie jwal...@biochem.wustl.edu) Subject: [ccp4bb] DNA oligonucleotide purification To: CCP4BB@JISCMAIL.AC.UK All, Our laboratory is looking for a new column to purify short (40mers or less) DNA oligonucleotides that will be used for crystallization setups. I wanted to ask if anyone has suggestions of a column that will give excellent separation of N and N-1 products. We have a Shimadzu HPLC system. We are considering Dionex columns as well as Oligonucleotide Separation C18 columns from Waters, but if there are others that are better we would like to know. Any suggestions will be greatly appreciated. Thank you! Jamie Wallen
