Why single colony?

"Lewis, Ben FMC Ben.Lewis at fmc.sa.gov.au
Thu Mar 13 20:23:46 EST 2003


Hi Edison,
Basically you need to create a master plate of one correct clone for further
use. The first plate after transformation can sometimes have irregular
clones. That is, not all colonies contain the desired insert. What can
happen is that if after incubation you receive a lot of colonies, the
antibiotic concentration in the plate can drop thus, allowing undesired
colonies to grow without the necessary vector. This is just a method to
ensure that every time you create a plasmid prep, you effectively have the
exact same clone. Hence, validating that data you collect is comparable.

Benjamin Lewis BSc (Hons) GRACI 

Research Associate
Department of Clinical Pharmacology,
Level 6, Flinders Medical Centre,
Flinders University School of Medicine,
Bedford Park,
South Australia 5042,
Australia. 

T  +61 8 82044795
F  +61 8 82045114
E ben.lewis at fmc.sa.gov.au
W www.flinders.sa.gov.au 



-----Original Message-----
From: chang_edison at hotmail.com [mailto:chang_edison at hotmail.com]
Sent: Thursday, March 13, 2003 10:11 PM
To: methods at hgmp.mrc.ac.uk
Subject: Why single colony?


After identifying the positive clones from the cell transformation
plates, why do we still need to streak and pick single colony for DNA
miniprep and protein? Don't all the decedents from that single
positive clone  have the same genetic made-up? What's the things we
try to avoid? Mutation in the cloned gene? Hetergeneous mixture of
plamids? What? Please be kind and blow up my super small brain. Thank
you for now! Cheers, EdiSon.
---



More information about the Methods mailing list