Media Fill of 96 well Inoculation plates |
Version Number: | 1 |
Start Production Date: | 2/05/00 |
Author: | Irmengard Rapier |
Edited by: | X |
Reviewed by: | Matt Fourcade |
Summary |
Materials & Reagents |
Materials/Reagents/Equipment | Vendor | Stock Number |
---|---|---|
Disposables | ||
96 well round bottomed Falcon plates | VRW | N/A |
Stock Solutions | ||
Terrific Broth with 1.6% Glycerol | Teknova | 0188-1L |
Ethanol | Aaper | N/A |
10X Phosphate Buffer | Teknova | 0190-1L |
Sterile H20 | Millipore | N/A |
Reagents | ||
Ampicillin | Sigma | N/A |
Equipment | ||
Titertek Stacker | Titertek | N/A |
Procedure |
1. Before using the Multidrop check that the machine has been calibrated on a weekly basis. Tubing should be changed on a monthly basis. Make sure the Multidrop has been properly lubricated and that the flexicable is in working condition.
2. Set the dispense mode to 185 ul and insure that the tubing is ready for use. Rinse the tubing with ethanol, priming for about 20 seconds. Let the ethanol sit in the tubing for approximately 5 minutes and then rinse thoroughly . Rinse with sterile ddH2O for about 20 seconds, making sure there is no ethanol left in the tubing.
3. Prime with fresh TB+amp for about 20 seconds and then begin filling plates.
4. Check the liquid levels in the plates and discard any that are uneven.
5. After use, repeat the ethanol and water rinses, leaving the tubing empty and wrapped in a clean paper towel.
6. Clean the area around the hood with ethanol after use.
Reagent/Stock Preparation |
How to prepare reagents and stocks:
1. Ampicillin (50mg/ml)
5 g Ampicillin
195 ml ddH20
Stir until dissolved. Refrigerate in a light proof container. This
has a 30-day expiration date.
2. TB/ampicillin
1 L Terrific Broth with 1.6% Glycerol
1 ml Ampicillin
100 ml 10x Phosphate Buffer
Mix thoroughly and refrigerate if not used immediately.