Chromosomal
DNA from E. coli cells or from a solid tissue.
Enough for 12 Isolations
Load isolated samples (optional) along with standard DNA
on a gel
Each gel can be shared by 2-3 students
Learn the process of DNA isolation from E.coli cells
in only 30 minute
protocol.
Learn how to make antibiotic resistant growth media.
Understand how high molecular weight genomic
DNA samples appear on a
gel as compared with
other samples of different sizes.
Learn how to stain, visualize and analyze DNA
samples on agarose gel
electrophoresis.
Isolation of genomic DNA is a major step in most cloning
experiments. E.coli cells are lysed by adding lysis solution
followed by brief incubation at 60º C. Most of the
proteins are separated after addition of protein precipitation
solution. After centrifugation, DNA solution (upper aqueous
phase) is removed, and DNA is precipitated with addition
of ethanol. DNA is resuspended in water for further experiments.
Isolated DNA can be loaded in a gel (agarose gel electrophoresis)
for visualization, quality and yield. DNA isolated by
this method can also be used for restriction enzyme digestion
or PCR (polymerase chain reaction) experiments.
Time Needed to Complete
the Experiment:
Grow E.coli cells (16 hours) or use frozen cells.
Lysis of E.coli cells: 10 minutes (about one hour if other
solid tissue is used) :
Precipitation of DNA: 15 minutes.
Run agarose gel electrophoresis (optional): 40 minutes.
Staining time:
2-3 hrs for methylene blue stain or instant results with
ethidium bromide stain.
Kit
includes following items:
E.coli cells (transformed with plasmid DNA)
Powder LB media for growth of E.coli cells (includes antibiotic
for E.Coli growth)
Protein precipitation solution
Standard DNA (100 µl) for comparing fragments of
various sizes.
Loading pipet of fix 20 µl volume. This pipet is
extremely durable which can be used up to
5,000 times.
Microfuge tubes rack of 96 loading tips.
Heavy duty test tube rack containing 80 holes for microfuge
tubes.
Agarose, enough for 10 gels
Concentrated electrophoresis buffer, enough for 10 gels
DNA staining solution.
Gel running buffer.
Measuring tubes (15 ml and 50 ml capacity) & microfuge
tubes.
Instruction sheet.
Blank sample for agarose gel as practice.
Videotape of Agarose Gel Electrophoresis.
Other
Requirements:
Beakers, Ethanol, Water and Test Tubes.
Catalog#
Description
Price(1
Kit)
Price(3
Kits)
Price(5 Kits)
306-1
Chromosomal DNA
from E.coli or solid tissue. 30 preps
$49.00
$139.00
$219
306-2
Extended Kit for
Chromosomal DNA from E.coli or solid tissue*
$84
$239
#389
*Extended
kit includes extra reagents (for visualization of
DNA during electrophoresis) such as loading pipet,
agarose, microfuge tube rack, gel buffer, standard
DNA, staining solution and agarose electrophoresis
videotape etc.
Enough for 12 Isolations
Learn the process of DNA isolation from E.coli cells
Grow E.coli cells (16 hours) or use frozen cells.
