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SOPs > E-0002 - HPC Pour Plate > Flashcards

E-0002 - HPC Pour Plate Flashcards

1
Q

What is the code for HPC?

A

SM 9215B

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2
Q

What is the first thing you do?

A

Make sure I have proper PPE on
- Lab coat, goggles, and gloves

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3
Q

What do you do after putting on PPE?

A

Turn off the AC and prepare the logbook

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4
Q

Preparing the logbook: what information do you include?

A
  • Prep date/time
  • Prep analyst initials
  • Media lot #
  • Pipette lot #
  • Balance ID
  • Plate lot ID #
  • Plate ID: “media”
  • Culture ID
  • Sample names and numbers under “comments”
  • DF (dilution factor): 1X (no dilution)
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5
Q

What do you do before setting up an HPC?

A

Turn off the AC

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6
Q

What do you do after turning off the AC?

A

Sterilize the work bench area, equipment, and gloves with 70% IPA

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7
Q

What do you do after sterilizing your work area?

A

Check the sample volume (100 mL)

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8
Q

What do you do after checking the sample volume?

A

Label the plates

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9
Q

What do you do after labeling the plates?

A

Open the air plate
- Record the air plate open date/time in the logbook

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10
Q

What do you do after opening the air plate?

A

Pipette 1.0 mL of sample onto each plate
- Shake the sample first

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11
Q

Describe the setup for an HPC.

A
  1. Line up sample bottles in order
  2. Line up air plate, media plate, and positive control in order
  3. Put two plates in front of each sample bottle (+2 for dups)
  4. Label each plate accordingly
  5. Open the air plate (record the air late open date/time)
  6. Have the media ready
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12
Q

What do you put on the batch sticker?

A
  • Day of week (abbreviation), batch setup alphanumeric letter for the day, analysis
  • Ex) Mon, A, HPC
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13
Q

What must you do before pipetting the samples on the plates?

A

Shake each sample 25 times

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14
Q

How much sample do you pipette onto each plate?

A

Pipette 1mL of sample onto the center of each plate

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15
Q

What media do you use?

A

HPC agar

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16
Q

What must you do before pipetting the media onto the plate?

A

Sterilize the rim of the bottle and the cap in the flame of a Bunsen burner

17
Q

How much media do you pipette onto each plate?

A

10mL

18
Q

What are things to keep in mind when pipetting the media?

A
  • Make sure the media isn’t too hot
  • Don’t pipette the media into the exact same spot as the sample
  • Add media to the positive control last
  • Swirl each plate as you go (don’t let the agar splash or touch the cover)
19
Q

What do you do after pipetting the media onto the plates with samples?

A

Prepare the QCs

20
Q

How do you prepare the QCs?

A
  1. Media plate
    - Pipette 10 mL of agar onto the plate
  2. Positive plate
    - Pipette 10 mL of agar onto the plate
    - Inoculate the agar with E. coli
  3. Air plate
    - You opened this before you started preparing your samples
    - After 15 minutes: add media and close the plate
21
Q

What do you do after preparing the QCs?

A

Weigh the media plate and record the weight in the logbook and on the chosen plate

22
Q

What do you do after weighing the media plate?

A

Add a batch sticker to the positive plate and the logbook

23
Q

What do you do after adding the batch sticker?

A

Incubate at 35 +/- 0.5 degrees C for 48 +/- 3 hours
- Record the incubator in time/temp
- Record the incubator ID

24
Q

What do you do after putting the samples in the incubator?

A

Calculate the read date/time
- Put post-it in the middle of the page

25
Q

Preparing the logbook for reading: what information do you include?

A
  • Read analyst initials
  • Read date/time
  • Incubator out time/temp
26
Q

After setting up the logbook for reading, what do you do?

A

Count the colonies on the QCs
- To make sure it fits the criteria at the bottom of the page

27
Q

What do you do after reading the QCs?

A

Count the colonies on the sample plates

28
Q

What do you do after counting the colonies on each sample plate?

A

Average the two counts obtained for each sample and dups
- Record in logbook under “Average Plate Count”
- Units: CFU

29
Q

How many significant figures do you report results in?

A

2 sig figs

30
Q

What do you do after reading the plates and recording the results?

A
  • Cross out the post-it and move it to the bottom of the page
  • Cross out the to-do list
31
Q

What do you do after crossing out the post-it and to-do list?

A

Put the samples and QCs on the 24-hour hold shelf

32
Q

List the general steps for an HPC (not including reading).

A
  1. PPE
  2. Get samples
  3. Logbook
  4. Turn off AC
  5. Sterilize area
  6. Pipette samples (1.0 mL)
  7. Open and sterilize media bottle
  8. Pipette media (10 mL)
  9. Prep QCs
  10. Weigh media plate
  11. Batch sticker
  12. Incubate
  13. Reading time

SOPs (8 decks)

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