Home  Contacts  Order  Catalog  Support
 Search  Alphabetical Index  Numerical Index
Fermentas logo
 Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification
 Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents
C E R T I F I C A T E
 

Microbiological Quality Control Procedure for FastMedia™ LB Agar

 Purpose

Test the sterility, selectivity and ability to support growth of a particular lot of selective FastMedia™ LB Agar and test the efficiency of colony color development of FastMedia™ LB Agar IPTG/X-Gal plates.

FastMedia™ being tested

 Positive control strain Negative control strain
LB Agar Amp, #M0021 JM109 pUC57 Amp JM109
LB Agar Amp IPTG/X-Gal, #M0031

JM109 pUC57 Amp

 JM109
 Material Needed for Each Pouch Tested
 Plate Preparation
  1. Pour the pouch contents into a 500 ml glass bottle or flask.
  2. Add 200 ml deionized or distilled water.
  3. Heat in a microwave oven on medium power until bubbles start to appear (2-3 minutes). Swirl gently to mix the preparation.
  4. Reheat the media for 30 seconds and swirl gently again. Do not over boil!
  5. Repeat step 4 if necessary until the media is completely dissolved.
  6. Allow the media to cool.
  7. Pour 8 plates.
  8. Have 4 plates with nothing streaked. Label these plates "I".
  9. Streak 2 plates with untransformed cells. Label these plates "II".
  10. Streak 2 plates with appropriate glycerol stock of positive control strain. Label these plates "III".
  11. Streak autoclaved conventional LB Agar plates with positive and negative control strains.  Label these plates "IV".
  12. Incubate all plates at 37°C.
 Plates Control
Plates Conditions Purpose Expected Result

I
 4 units

 Unstreaked Control for sterility of equipment and procedure No growth

II
2 units

 Streaked with negative control strain Control for antibiotic presence in the media and no growth of negative control strain No growth
III
 2 units		 Streaked with positive control strain Control for growth support of positive control strain Growth
IV
 2 units		 Autoclaved conventional LB media streaked with positive and negative control strains Control for viability of positive and negative control strains Growth
 Results

After 16, 24 and 48h incubation, check each group of plates for growth and blue color development and record in chart below:
(+) growth,
(-) no growth.

Plates Result
after 16 hours after 24 hours after 48 hours
I      

II

      
III      
IV      

Blue after 16 hours ______  (IPTG/X-Gal media only)

Results should indicate all plates "III", plates "IV" as having growth and all plates "I" as having no growth. The plates "II" should also have no growth. When testing the IPTG/X-Gal media plates, blue colonies should be visible after 16 hours of incubation at 37°C. Results are measured by visual inspection. If results agree with these indications, the lot passes. If there is growth on plates "I" or plates "II", the lot fails. Additionally, if there is no growth on any of the plates "III", or if blue colonies are not visible on IPTG/X-Gal plates, the lot also fails. Repeat all tests if lot fails.

If lot fails "QC", repeat the QC test in it's entirety.

If lot fails "QC" again, it's identified, segregated and destroyed.

 Home  Search  Contacts  Order  Catalog  Support

Updated gegužės 10, 2007 09:40