"
Team:Newcastle/Gel extraction
From 2010.igem.org
(Difference between revisions)
RachelBoyd (Talk | contribs) (New page: ====QIAquick gel extraction microcentrifuge protocol==== # Excise the DNA fragment from the agarose gel with a clean, sharp scalpel # Weight the gel slice and add 3 volumes of buffer QG to...) |
RachelBoyd (Talk | contribs) (→QIAquick gel extraction microcentrifuge protocol) |
||
| Line 1: | Line 1: | ||
| + | {{Team:Newcastle/mainbanner}} | ||
| + | |||
====QIAquick gel extraction microcentrifuge protocol==== | ====QIAquick gel extraction microcentrifuge protocol==== | ||
# Excise the DNA fragment from the agarose gel with a clean, sharp scalpel | # Excise the DNA fragment from the agarose gel with a clean, sharp scalpel | ||
| Line 14: | Line 16: | ||
# To elute DNA, add 30 µl of Buffer EB to the center of the QIAquick membrane and allow to stand for 1 min | # To elute DNA, add 30 µl of Buffer EB to the center of the QIAquick membrane and allow to stand for 1 min | ||
# Centrifuge the column for 1 min and transfer the eluate to a clean 1.5 ml micriocentrifuge tube | # Centrifuge the column for 1 min and transfer the eluate to a clean 1.5 ml micriocentrifuge tube | ||
| + | |||
| + | {{Team:Newcastle/footer}} | ||
Revision as of 08:13, 10 August 2010
| |||||||||||||
| |||||||||||||
QIAquick gel extraction microcentrifuge protocol
- Excise the DNA fragment from the agarose gel with a clean, sharp scalpel
- Weight the gel slice and add 3 volumes of buffer QG to 1 volume of gel (100 mg ~ 100 µl)
- Incubate at 50°C and invert the tube gently at regular intrerval until the gel has completely dissolved
- After the gel has dissovled completely, check that the color of the mixture is yellow
- Add 1 gel volume of isopropanol to the sample and mix
- Place a QIAquick spin column in a 2 ml collection tube
- To bind DNA, apple the sample to the QIAquick column and centrifuge for 1 min
- Discard the flow through and place the QIAquick column back into the same tube (Maximum volumn is 750ul)
- Add 0.5 ml of Buffer QG to QIAquick column and centrifuge for 1 min and discard the flow through and place the QIAquick column back into the same tube
- To wash, add 0.75 ml of Buffer PE to QIAquick column and centrifuge for 1 min and place the QIAquick column back into the same tube
- Centrifuge the column for a further 1 min
- Transfer the column into a clean 1.5 ml micriocentrifuge tube
- To elute DNA, add 30 µl of Buffer EB to the center of the QIAquick membrane and allow to stand for 1 min
- Centrifuge the column for 1 min and transfer the eluate to a clean 1.5 ml micriocentrifuge tube
   
|
    
|
