Barney:PCR purification

Materials
Qiagen QiaQuick Spin columns Qiagen Buffer PB
 * OR
 * 5M Guanidine Hydrochloride
 * 10mM Tris-HCl pH 6.6
 * 30% ethanol

Qiagen Buffer PE
 * OR
 * 10mM Tris-HCl pH 7.5
 * 70% ethanol

Qiagen Buffer EB
 * OR
 * 10mM Tris-HCl pH 8.5

DNAse-free Milli-Q Water for elution (should be slightly basic, pH 7.0-8.5) 1M HCl

Protocol
|Qiagen kit manual
 * 1) Bind DNA
 * 2) Add 5 volumes of Buffer PB (or equivalent) to PCR reaction to be purified (i.e. add 250μL to a 50μL PCR reaction)
 * 3) Load onto a clean spin column, place the column in a clean collection tube, cap, and centrifuge (5000rpm, 1 minute)
 * 4) Discard flow-through, place column back in empty collection tube
 * 5) Wash
 * 6) Add 700μL of Buffer PE (or equivalent) to column and centrifuge (13000rpm, 1 minute)
 * 7) Discard flow-through
 * 8) Replace empty column in collection tube and centrifuge again (13000rpm, 1 minute) to dry column
 * 9) Elution for Sequencing
 * 10) add 30-50μL of DNAse-free Milli-Q water to the center of the column
 * 11) incubate for 1 minute at room temperature
 * 12) place column into new, sterile DNAse-free tube for DNA collection
 * 13) centrifuge (13000rpm, 1 minute)
 * 14) Elution for downstream processing
 * 15) add 30-50μL of Buffer EB to the center of the column
 * 16) incubate for 1 minute at room temperature
 * 17) place column into new, sterile DNAse-free tube for DNA collection
 * 18) centrifuge (13000rpm, 1 minute)

Column Re-Use Protocol

 * 1) step 1
 * 2) step 2
 * 3) step 3
 * 4) etc.