Storage procedure for yeast preservation (SOP ID: 0011)

SOP ID:

0011

Revision nr.:

0.0

Release date:

January 28, 2020

Last release date:

January 28, 2020

Author(s):

Ada Serena Marletta

Approver:

Ada Serena Marletta

1. Scope

This Standard Operating Procedure provides a step-by-step protocol to prepare yeast strain glycerol stocks.

2. Background

This Standard Operating Procedure is intended for qualified personnel who was trained according to the ISO-9001 quality framework.

3. Prerequisites

Documents required:

MIAMI quality manual
SOP ID 0001

Material required:

Generic lab-, glass- and plastic-ware
30% Sterile glycerol solution
Sterile YPD medium
2 ml screw-cap vials
dry ice
orbital shaker
spectrophotometer

4. Changes since last revision

No changes were made since last version

5. Role and responsibilities

Author (Au): Staff scientist properly trained to draft, review and disseminate standard operating procedures within the framework of ISO-9001

Approver (Ap): PI with the authority to approve, reject and withdraw standard operating procedures

MIAMI Quality Manager (QM): MIAMI authorized Quality Manager

Operator (Op): Staff scientist properly trained to execute the standard operating procedure

6. Monitoring Requirements

The present document can be reviewed at any time required by the author and the MIAMI Quality Manager.

7. Procedure

Step	Action	Responsibility
1	Take two 14 ml tubes, one tube for the sample and one tube for the negative control, and mark them with “sample name” and “ctrl-”, respectively.	Op
2	Add 5 ml of sterile YPD to each 14 ml tube.	Op
3	From a plate, pick a single colony of the yeast strain and inoculate the “sample name” tube.	Op
4	Put the “sample name” and the ctrl-” tubes in the orbital shaker diagonally and incubate overnight at 30°C, shaking at 250 rpm.	Op
5	After the overnight growth, the “ctrl-” tube should be clear and the “sample name” tube should appear turbid. IF Both conditions are satisfied, THEN proceed to step 7 IF the “ctrl-” tube appears turbid, THEN go to step 6 IF the”sample name” tube appears clear, then go to step 1	Op
6	When the “ctrl-” tube is turbid, it means that the culture medium was contaminated. Take a new sterile YPD medium and go back to step 1	Op
7	Measure the OD600 of the “sample name” tube: IF OD600 is >2, then proceed to step 8 ELSE keep growing at the same conditions reported at step 4 until OD600 >2.	Op
8	Take two sterile 2mL screw-cap vials. Write the stock number on the top and the strain genotype on the side	Op
9	Pipet 0.5 ml of the 30% Glycerol (w/v) solution into each of the sterile 2mL screw-cap vials.	Op
10	To each vial, add 0.5 mL of the stationary phase culture, mix, and screw caps.	Op
11	Add one vial to the appropriate yeast stock box in the main -80°C and the other vial to the backup stock box.	Op

8. Record Management

When completed and approved, the SOP will be published on MIAMI website

9. References

9.1 Standards

ISO-9001:2015
International System of Units

9.2 Other SOPs

The execution of this SOP does not refer to any other SOP

9.3 Reagents

For reference reagents, please refer to the Prerequisites section.

9.4 Sequences

The execution of this SOP does not refer to any reference sequence

9.5 Supplementary Documents

There are no supplementary documents.

10. Definitions

Measurements units are defined according to International System of Units

Metric prefixes are defined according to Metric (SI) Prefixes

Whenever possible, biological construct are defined according to the SBOL standard

Whenever possible, biological experiments are defined according to the metrology standards

11. Disclaimers

This Standard Operating Procedure (SOP) was developed based on a compilation of best available information, knowledge, experience, and general industry practices to provide guidance to MIAMI staff in performing the activities defined herein, in a consistent and standardized manner. This document does not contain regulatory or statutory requirements unless specified. The MIAMI consortium has made every attempt to present the information in a clear and concise manner for a variety of users. However, the MIAMI consortium is not responsible for the misuse or misinterpretation of the information presented herein. Under no circumstances shall The MIAMI consortium be liable for any actions taken or omissions made by non-MIAMI users of this document. In general, this document should be used as a reference. Differences may exist between the procedures referenced in this document and what is appropriate under project-specific conditions. This document does not represent an endorsement of practitioners or products mentioned in the document.

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