Diluting and Freezing Primers
- Grab the primer binder located on the computer desk and open it up.
- Open up the Dropbox folder on the computer and proceed to the folder named “mccormicklabshared”.
- Within the folder locate the MM-Primer sheet and open it. It should be showing as soon as you open up the “mccormicklabshared” folder.
- Take the Packing List sheet that was given alongside the order of primers. On it, write down what MM numbers the new primers should encompass. For example, if the latest primer in the binder was MM159 and your packing list includes two primers, you should write down MM160-161.
- On the specification sheet of each primer write down what its MM number will be at the top in huge writing.
- Once the primer has an MM number you must highlight the sequence name, the sequence of base pairs, the amount of nanomoles, and the amount of water to add to the primer.
- After highlighting the appropriate information on the specification sheet, write down how much water should be added to the primer. If the amount of base pairs is known or specified to you, then also write down that information on the specification sheet.
- On the MM-Primer sheet that should be open on the desktop, you should copy the appropriate information from the specification sheets into each column. *For the sequence and name information, someone should either have it in an excel spreadsheet or email if they ordered it. Copy and paste this information into their appropriate columns to avoid putting down the wrong information.
- When finished copying down information into the MM-Primers spreadsheet and saving it, place stickers onto the primers that correspond to their MM numbers. Always check that each primer has the correct MM number and that the MM number is paired with the correct primer, as it helps ensure everything is labeled correctly.
- Once done labeling each primer, centrifuge the primer tube(s) to ensure all DNA is at the bottom of each primer tube.
- Taking your specification sheet with you, specify how much water must be added to the correct primer tube and pipette that amount to the primer tube. Always check that the amount of water you are adding corresponds to the correct amount noted on the specification sheet and you are adding the right amount of water to the appropriate primer tube. When adding water to multiple primer tubes, always replace pipette tips to avoid contamination.
- When finished pipetting water to the primer tube(s), vortex them to ensure that all DNA has been mixed within the solution. To make sure of this, check the bottom of the primer tube to see if any DNA is still settled.
- Once all DNA has been completely mixed within the solution, place the primer tube into the appropriate primer tube box within the -20C Freezer.
- When finished with the specification sheets and their packing list, hole punch them and place them in the primer binder after the latest MM numbered primer.