Class Notes (810,968)
Canada (494,410)
Biology (Sci) (2,418)
BIOL 301 (19)

BIOL 301 Lab 8.docx

2 Pages
Unlock Document

McGill University
Biology (Sci)
BIOL 301
Nam Sung Moon

BIOL 301 Lab Muhammad Fazal 260416908 1. Preparing a Total Extract  Add 200 ul of Lysis Buffer to the tube containing the yeast pellet and vortex until cells are completely resuspended in the buffer.  Pour all the beads (200 ul) into the previous suspension  Vortex the tube for 1 minute at high  Place on ice for 1 minute. (Always keep on ice from now on)  Repeat steps 3 and 4, 7 times  Label, place them in the centrifuge  Spin for 5 minutes at 2000 rpm at 4C  Transfer supernatant (200 ul) to a new tube.  Add 6 ul RNase/DNase to the supernatant  Vortex at 5 for 30 seconds  Incubate on ice for 10 minutes  Add 300 ul of Extraction Buffer and invert the tubes 4-5 times (avoid foam)  Take tubes on ice to 4C fridge and clamp them in the rotator for 30 minutes  Place the labeled tubes in the refrigerated centrifuge  Spin at 4C for 10 minutes at 12000 rpm  Transfer the supernatant (400 ul) to a micro tube marked TOTAL and place on ice.  From each TOTAL extract transfer 10 ul into a new micro tube and keep on ice until Part 3  Add 20 ul from the TOTAL to a new micro tube with 2 ul of Storage Buffer. Label properly. Mix and put immediately on bucket labeled dry ice. 2. Affinity Purification  Transfer 370 ul of TOTAL from Part 1 to the tube containing IgG and label  Mix by inv
More Less

Related notes for BIOL 301

Log In


Don't have an account?

Join OneClass

Access over 10 million pages of study
documents for 1.3 million courses.

Sign up

Join to view


By registering, I agree to the Terms and Privacy Policies
Already have an account?
Just a few more details

So we can recommend you notes for your school.

Reset Password

Please enter below the email address you registered with and we will send you a link to reset your password.

Add your courses

Get notes from the top students in your class.