Appendix: Radiation Safety Procedures

June 8, 1990

Matthew S. Holt, RSO

General Rules

1. Review the WUMS Radiation Safety Manual, especially the following sections:
   • Section 4. Regulations
   • Section 5. "ALARA" (as low as reasonably achievable) program
   • Section 6. Training program for personnel working with RAM (radioactive materials)
   • Section 7. Responsibilities of individuals working with RAM
   • Section 10. Instructions for receiving and opening RAM packages
   • Section 11. General rules for safe handling of RAM
   • Section 12. Personnel Bioassay program
   • Section 13. Information regarding Biological effects of Radiation exposure
   • Section 15. Instructions regarding Prenatal Radiation exposure
   • Section 17. Instructions regarding accidents involving RAM
   • Section 20. Information regarding services provided by the Radiation Safety staff

   • Review the training manual (see RSO) for information pertaining to passing the radiation safety examination. All personnel must pass this exam (or a comparable one from another institution) prior to working with RAM.
   • All RAM users are required to sign a contract with Washington University agreeing to comply with all rules and regulations.
   • Account for your isotope as you use it.
   • Record all drain disposals promptly.
   • Monitor before, during, and after each use. Change gloves when necessary.
   • Understand and practice ALARA principles.

1. Plan experiments and have everything ready before using the isotope. Record time for use of the radioactive center. Be prepared to accommodate others when necessary.
2. Monitor before, during, and after each use of isotope. Monitor gloves frequently. Low counts can be rinsed off gloves into radioactive disposal sink.
3. All RAM users must leave the center clean. If you find the workstation contaminated before starting, it is your responsibility to correct the problem by either informing the person responsible or cleaning the area yourself.
4. Use 50 ml conical tubes to contain the used columns (when dry, the column resin Biogel P-60, can pose a severe health risk to those handling the radioactive waste).
5. Work as quickly as possible to minimize your exposure, but at a pace with which you are comfortable.
6. Pack radioactive waste into proper waste containers. Fill one keg before starting the next.
7. Contain and immediately report any accidents or spills to the Radiation Safety Officer, (Matthew S. Holt), or Andrea Burgess if Matt is unavailable. Inform others of potential hazards and/or procedures that may result in exposure.

Waste Disposal

1. Obtain a Radioactive Waste Transfer Form from the W.U. Radiation Safety Office (362-3475), from the waste disposal truck driver, or from the RSO for the lab, Matthew S. Holt.
2. Waste pick-up is limited to Monday, Wednesday, and Friday from 9:00 to 9:30.
3. Dry waste keg sizes are 1.1 cu. ft. and 0.65 cu. ft. Pack as tightly as possible. Different isotopes must be segregated and each keg must have a transfer form. Half kegs can be disposed and should be marked accordingly.
4. Liquid waste keg sizes are 5 gals and 2 gals. Different isotopes must be segregated and each keg must have a transfer form. Estimate the volume for partially filled kegs and mark accordingly for disposal.
5. Activity must be calculated for each waste container. Dry waste activity is calculated by taking 2/3 of the mCurie order total and dividing it by the number of dry waste kegs for the week. Liquid waste activity is calculated by taking 1/3 of the mCurie total and dividing it by the number of liquid waste kegs for the week. A 1 ml aliquot of the liquid waste is then counted to verify the computation. Activity should be reported in microCuries (e.g., < 750 uCi).
6. Wipe tests of all waste kegs must be performed and the dpm/100 cm[2] must be recorded on the waste transfer form.
7. Meter survey results must be determined in mR/hr on contact and at 1 meter and recorded on the waste transfer form.

Radioactive Sink Disposals

Since each hybridization contains a different amount of radioactivity, the number of dpm in each wash will vary. Do not dispose of a liquid down the radioactive drain until it has been monitored, counted in an LSC, and determined to be of a sufficiently low level of radiation. The geiger counter can assist in determining whether a wash can go down the drain. If the wash solution of 500 ml has a meter reading of less than 1 mR/hr, take an aliquot of the solution and count it in an LSC. Enter the total counts on the drain disposal record. Generally speaking, the first "Postwash 2" solution has a meter reading of less than 1 mR/hr (this is not true when hybridizing more than 10 blots). Defining 1 mR/hr as 4 x 10e4 cpm, a 1 mR/hr reading of 500 ml approximates 4 x 10e4 counts for [32]P. One uCi equals 2.2 x 10e6 counts, hence a 1 mR/hr reading represents approximately 0.02 uCi of RAM.

Wipe tests

1. Wipe tests are performed weekly for all areas in which radioactive materials are used or stored as well as on each keg for disposal. Blank wipe test forms are available from the RSO or the W.U. Radiation Safety Office. Wipe test results are filed in the Radiation Accountability Notebook maintained by the RSO.

2. Any area with "dpms" higher than 200 must be cleaned. After cleaning, perform another wipe test for documentation.

3. Use Wheaton Omni-vials (cat.#225402) and the Beckman LS 3801 Scintillator on "User Number 2" for e32P dpms.

Receiving Isotope

Each package containing radioactive materials has a packing slip and a Radioactive Shipment Receipt Report (which must be completed by lab personnel) for all the isotope contained in that package. Check the packing slip and vials to verify the radionucleotide, activity, and chemical form. Remove the RAM and survey the shipping carton and packing material. Wipe the innermost container (vials) with an EtOH soaked kimwipe and hold to a monitor to check for contamination. We are required to perform a wipe test on the packing material and the shipping carton for Tritium shipments only. Dispose of packing materials and shipping cartons in the regular trash after marking out all radiation labels.

RSO Duties

1. Maintain the lab Radiation Accountability notebook consisting of employee contracts and the review session attendance, isotope consumption (record for each use of isotope), shipping package documentation, and weekly wipe tests.
2. Calibrate all lab geiger counters yearly and file the documentation.
3. Organize and present yearly review sessions for all users of RAM.
4. Order isotope.
5. Dispose of waste.
6. Check in isotope.

Radioactive Label, all 10 mCi/ml	Amersham Cat #	Primary use in lab
alpha-e32P-dATP, 3000Ci/mmol	PB10204	primer extension or nick translation
alpha-e32P-dCTP, 3000Ci/mmol	PB10205	primer extension or nick translation
alpha-e32P-dTTP, 3000Ci/mmol	PB10207	nick translation and PCR
alpha-e32P -ATP, 800Ci/mmol	PB10384	PCR or sequencing
gamma-e32P-dATP, 3000Ci/mmol	PB10218	kinase (end labeling)
alpha-e35S- dATP, 3000Ci/mmoll	SJ1304	sequencing

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