Dna Gels - Thermo Scientific OWL HEP-1 Operating And Maintenance Manual

DNA Gels

Thermo Scientific
Preparing a sequencing gel for transfer on the HEP-3
1. Remove the gel cassette from the gel box and place under cold,
running tap water until the surfaces of both glass plates are cool. This
facilitates handling of the gel and prevents the gel from "curling" as it
cools once it is removed from between the glass plates.
2. Place the glass plate sandwich, with the notched or shorter plate facing
up, flat on paper towels on the lab bench and remove any excess liquid
and remaining tape (or binder clips).
3. Remove one side spacer (wearing protective gloves) and insert a long
metal spatula between the glass plates where the spacer had been
present. Gently rock the spatula, forcing separation of the plate from
the gel. The gel will normally remain affixed to the bottom (not
notched) plate. Remove the top (notched) plate by slowly lifting it
from the side with the inserted spatula and gradually increasing the
angle lmtil the top plate is completely separated form the gel. If the gel
sticks to the top plate in an isolated spot, a stream of water from a
wash bottle can be sprayed at the spot to aid separation. Occasionally,
a gel sticks to the top plate instead of the bottom plate. In this case,
flip the gel sandwich over and follow the same procedure.
4. Once the plates are separated, remove the second side spacer along
with any extraneous bits of acrylamide around the gel. If excess water
was utilized to aid in the separation of the gel from the glass plate, use
a paper towel to absorb the excess liquid. Using a piece of dry blotting
paper(Whatman 3MM chromatography paper, 46 x 57cm), gently roll
it onto the gel beginning at one end and working slowly towards the
other end. Care should be taken to prevent air bubbles from forming
between the paper and the gel. Once the blotting paper is in position,
set another glass plate on top of the paper. Flip the entire sandwich
over. The gel should now be resting on top of the filter paper between
the two pieces of glass. Position the sandwich with one long side of the
glass hanging about one-third of the way off of the counter top. Slowly
push the bottom plate back toward the counter top, holding the top
plate in place, which will allow the blotting paper and gel to peel away
from the top piece of glass. The weight of the gel and blotting paper
causes the gel to slowly peel away from the top plate. Continue this
process until the gel is almost entirely peeled away from the glass. A
small portion of the gel may still be attached to the glass; in this case, a
small stream of water from a wash bottle can be used to aid in removal
of the gel.
Section 2
Setting Up
Semidry Electroblotter 2-3