Crandall Lab

1.    Obtain Millipore multiscreen filter plates (96 well).  

2.    Pour Sephadex on plate loader and fill all holes.  Place left over sephadex back into bottle.

3.    Turn filter plate upside down and slide onto plate loader.  Tap plate loader to get the sephadex to fall into the wells.

4.    Add 300ul dH2O to each well  (make sure the wells are full)

5.    Add a used v-bottom 96well plate to the bottom of filter plate making sure rows A-H are aligned on both plates

6.    Let plate stand for 15 minutes

7.    Spin plate in centrifuge (using balance plate with water) for 3 minutes at 3700rpm.  

8.    Empty and rinse v-bottom 96 well plate and place back in cupboard.

9.    Add a new (labeled) v-bottom 96 well plate to filter plate making sure rows A-H are aligned on both plates.

10.    Add 15ul dH2O to sequencing reaction

11.    Transfer all sequencing samples to filter plate

12.    Spin plate for 3 minutes at 3700 rpm.

a.    Dump out used columns and rinse filter plate thoroughly with dH2O and let dry.  Return balance plate to fridge.

13.    Dry samples in Vacufuge at 60º for 30 minutes.  Make sure to use balance plate

14.    Cover plate and mark off all unused wells.

15.    Write DNA sequence submission number on plate and cover

16.    Place plate in DNASC fridge.