Template preparation and purification

eagle-i ID

Resource Type

Properties

1. Fee for service

   Yes

2. Resource Description

   "Template DNA can be prepared and/or purified by the facility staff. Plasmid, cosmid, lambda phage, and BAC preps are offered through the Sequencing Facility. PCR reactions and purification using commercial columns is also offered by the facility. Plasmid minipreps are done on the Qiagen BioRobot. These preps yield 5-10 µg DNA for high copy number plasmids. If used for sequencing, the Facility guarantees the results, i.e. we will repurify and repeat sequencing at no additional charge until successful sequence is obtained. The facility also offers reliable treatment of marginal or bad templates to make them suitable for automated sequencing. This involves RNase treatment followed by column purification or phenol-chloroform extraction. This treatment improves quality of template, producing more reliable sequence and longer reads. For primer walking projects we recommend that a substantial amount of DNA (nominally 4-5 µg per kb for each direction) be treated this way and be used for the whole project. Templates prepared or purified by the facility staff are guaranteed to produce good sequence with standard, reliable primers, barring sequence specific problems (GC rich templates, secondary structure, internal repeats). The facility also provides free training in template preparation and troubleshooting, at users' request."

3. Service Fee URL

   http://www.med.upenn.edu/genetics/dnaseq/charges.shtml

4. Service Provided by

   DNA Sequencing Facility (Penn)

5. Website(s)

   http://www.med.upenn.edu/genetics/dnaseq/plasmid.shtml

6. Related Technique

   DNA purification

7. Related Technique

   Plasmid purification

8. Related Technique

   DNA sequencing assay