Materials

• Samples to be sequenced
• PCR tube (for one samples) or PCR strip tubes (for multiple samples)
• Parafilm
• 15 mL conical tube

Protocol

1. Quantify the concentration of DNA in each sample using the Qubit BR dsDNA kit.
2. Dilute sample so that there is 30 ng/µL concentration with a volume of 10 uL into a PCR tube.
1. You need 300 ng of DNA in the 10 µL so you need to find the volume needed from the sample.
1. E.g., Qubit output = 42.6 ng/µL to find the volume containing 300 ng you divide 300n g by the concentration and will get the volume of sample needed = 7.04 µL
2. The remaining volume to get to 10 µL should be water.
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For one sample, the DNA should be diluted in a single PCR tube. For multiple samples, you should use a PCR strip tube.
3. Ask Dr. Wheeler to submit the sequencing order on the Plasmidsaurus website.
4. Label the tube (or first tube of the strip tube) with the 3-character code indicated on the receipt.
1. If you are shipping more than one tube, label the first tube with the 3-character code and on the second tube write the strip number (if submitting multiple strips).
2. Do not add anything else to the labels.


5. Package the samples by placing a piece of Parafilm around the tubes to ensure that they do not open during shipment. Then put the samples into a protective box.
1. If one sample, place it in a 15 mL tube.
2. If multiple samples use a small box.


6. Ask Dr. Wheeler to prepare a FedEx shipping label and fill out the mail services eForm to request a pickup.
7. Place the prepared samples in a FedEx envelope with the shipping label and leave the biology office for pickup by mail services.