Day 3: Time for inoculation and sampling

Inoculation:

    For the day 3,  we have done our inoculation using minifors bioreactor. But we have to consider the proper steps to ensure our inoculation nicely done. Remember, all the steps must be followed!!

How to start inoculation?

• First, you have to sterilize your hands which have been covering with gloves with 70% ethanol. REMEMBER: it necessary to keep in sterilizes condition. 

• Before we continue our inoculation, the bioreactor must be set up at 700 rpm (speed of agitator). Then, the injections of nitrogen gas are done to remove oxygen in the bioreactor. 

• After that, we spray the inoculation tube and inoculation flask with 70% ethanol. The clamp and silicone tube are opened. 

• Remove the foil cover from flask and the inoculums were pour quickly into the vessels of bioreactors. 

• The port closure was placed onto the vessel top plate to cover the hole dan screw it down until hand tight. 

• The process was started.

 SAMPLING

Taking a sample without letting contaminants get into the vessel is simple if we follow the steps below:

1.   The sample device will normally be closed off from the vessel by  clamp on the silicone tubing leading to a pipe in the vessel

2.   The clamp is removed

3.   A syringe is fitted to the air filter connected to the short pipe on the head plate of the sample device

4.   Pull back on the syringe plunger to draw culture into the sample bottle

5.   When the sample has reached 2/3 full of the bottle, push down on the syringe plunger to return culture int the tubing into the vessel.

6.   Clamp the tubing closed again and the sample was take out by syringe.to the empty tube

    The reading of the sample ware taken every two hours starting from 12.50 p.m to 10.50 a.m. This sampling activity done almost 12 hours. The optical density of the sample was measured via Spectrophotometer at 600nm and glucose level was measured via glucose detector. Based on the results, the graph have been constructed.