The SOP • Technicians working in a biotechnology manufacturing facility must follow procedures exactly every time • Companies, therefore, produce detailed step-by-step instructions called a “Standard Operating Procedure”, or SOP • Is a very formal set of instructions, including precautions and lists of necessary materials. • Well-written SOP’s provide direction, improve communication, reduce training time and improve work consistency • Forces a person to think through a procedure step by step - it becomes a useful training took and reminder of GLP’s • Every company has its own criteria for writing SOP’s, and some are very detailed
Topics That Lend Themselves to SOP Format • Lab safety • Disposal of hazardous materials • Surface Decontamination • Spill Procedure • Inventory Procedures • Operation and maintenance of equipment • example: how to run a PCR reaction • using a spectrophotometer
Freestall MaintenanceEffective date: 10/1/2001Developed by: Milking staff • Key Objectives: • Clean stalls to create a sanitary bed for cows to lie on • Distribute bedding evenly to promote cow comfort • Notes: • Freestall rake is needed for this job • Freestall maintenance should be completed immediately after a group of cows is moved to the holding area • The Barn Operator is responsible for freestall maintenance • Scrape any manure or wet bedding from the freestall beds into the alley. • Rake bedding from front of the freestall beds toward the back. • Evenly distribute the bedding so that at least 1 inch of material covers all parts of the bed. • Observe free stall beds, divider loops, and other parts of the barn for any foreign objects, loose bolts, or other unusual conditions. • Return freestall rake to storage area at end of barn. • Write any unusual observations or repairs needed in the milking shift notebook. EXAMPLE - AGRICULTURAL
EXAMPLE 2 - PCR Protocol • Standard Protocol: 55ul Total PCR Reaction Volume: • Using gloved hands, remove the master mixes from the freezer. Allow the tubes to thaw; then gently vortex to mix. • In a containment hood or dead air box remove an appropriate aliquot to clean, sterile microfuge tube (one tube for each of the master mixes). Aliquot volumes should be 50ul for each sample + 150ul for the positive, negative and no template controls. We recommend adding an additional 20ul to correct for pipetting errors. • Add the appropriate amount of either AmpliTaq Gold or AmpliTaq DNA polymerase (0.25ul of either AmpliTaq Gold or AmpliTaq @ 5U/ul per 55ul total PCR reaction volume) to each of the master mixes and gently mix by inverting several times or gently vortexing. • Aliquot 50ul of master mix to individual wells of a PCR plate. • Add 5ul of DNA from the unknown and control samples to individual tubes or wells containing the respective master mix reactions, and pipette up and down several times to mix. Amplify target DNA using the universal thermocycler program. Continued on next slide
Specimen Requirements • This assay tests genomic DNA or complementary DNA (cDNA) from the following sources: • 5cc of peripheral blood, bone marrow biopsy, or bone marrow aspirate anti-coagulated with heparin or EDTA • Minimum 5mm cube of tissue • 2ug of genomic DNA • 5ug of total RNA or mRNA • 1ug of cDNA • Formalin-fixed paraffin embedded tissue or slides
SOP Writing Activity • We will form 10 groups and each will be responsible for writing an SOP for a common activity. • Each group will present there SOP to the class for constructive criticism.
Groups • Ricky & Emily • Alex ♂ & Lindsey • Kristen & Yuriy • Lauren & Devin • Zach & Liz • Amanda & Kat • Avelina & Ryan • Dylan & Alex ♀ • Andrew & Allie • Rawad & Scott & Megan
List of SOPs • Making a PBJ sandwich • Making Mac and Cheese • Taking a shower • Changing a flat tire • Coming to school in the morning • Solve the following algebraic equation: (3y + 7)/5 = (2y-12)/3 • Loading dishes in the dishwasher • Walk from here to the front of the building • Properly arrive at an airport and board a plane • Build a fire and make s’mores