This is going to be a very lab-specific instructional post, since it’s literally only going to talk about how we normally order primers here in the lab.
So as I mentioned in this previous post, at least for us here at CWRU, primers are cheapest when ordered from ThermoFisher. There’s an unfortunate delay on how long you have to wait until you can receive the primers, but for the most part it isn’t a big problem. Regardless, I’ve essentially streamlined how we order and organize primers as much as possible, so here are the steps.
Any permanent member of the lab should have their own tab (titled with their initials) in the “MatreyekLab_Primer_Inventory” sheet. I also have my own tab, which is called “KAM”. Every primer you order should have its own unique identifier (like KAM2199). Since you’re the only one adding to your own tab, there is no reason you should ever have duplicate identifiers. You want to do this step first, so you know what identifier to give the primer you intend on ordering now (The most recent primer I ordered was KAM3495, so the primer I’ll order today is KAM3496).
I usually keep a separate “scratch” google sheet where I write down things I’m in the process of doing. Here, I like to make a set of fields like this (I usually just copy-paste from the last time I ordered primers, actually). But, if you don’t already have one of these sheets already, I just made a “Scratch_area” tab in the google sheet.
Here, I like to list a few things. There’s an area I intend to later copy-paste into the primer inventory, which lists the date, the length of the oligo (using the =LEN() function of the actual primer sequence), the primer identifier, the actual primer sequence, and then a short description for what the primer is for.
Next, I like to have another section which repeats a subset of the information, but in a slightly different order. The reason for this is that the ThermoFisher website requires the primer ordering sample sheets to have the data entered in a specific order (sequence, identifier, Name of person ordering). Since this is the same information as in some of the previous columns, I just have those cells point to each of the relevant cells so that I don’t need to fill in that same info again.
If you’re ordering primers for sequencing, then you’re all set. But if you’re ordering primers for some cloning, I like to also write a short section that writes down which primers are supposed to be paired with which other primers to make a particular plasmid, also listing the template plasmid that will be used for the reaction. It’s nice to write this info now, so you don’t have to try to remember everything / redo the effort you just did to remember how you need to pair things to actually perform the PCR when you receive the primer. I usually copy-paste these cells into the “MatreyekLab_Hifi_Reactions” google sheet, into an area below where I have written “** denotes primers we may not have yet“.
So next is actually ordering the primers. There is a folder in the lab google drive where all of the primer order sheets are kept (MatreyekLab_GoogleDrive/Primer_order_templates). I usually just find a recent file, duplicate it, and rename it for today’s date and whoever is ordering (eg. KAM).
Once I copy-paste the new info in, it should look like this in the end.
K, great. Now to actually order it. Log into our thermofisher account by going to www.thermofisher.com, hitting “Sign In” at the top right. Lab account email is “[email protected]” and the password is the lab password (which you should already have memorized!). Then I hover over “Popular” at the top left, and click on “Oligos, Primers, & Probes”.
Then hit “Order now” under “Custom made to order”
Then I always do the “Bulk Upload” button:
Next hit “choose file”:
Then click on the samplesheet you want to upload. Your primer should now be entered.
Cool, so now is actually checking out. Hover over the cart and then click ‘View cart & check out”.
Next is a series of somewhat annoying confirmation screens. Scroll down and hit “Begin Checkout”
Default settings are fine, so scroll down and hit “Continue to Payment”
Default settings (including the speed type) are still fine, so scroll down and hit “Continue to Review Order”
You’re almost done! Now scroll down, click on the “You agree to the thermofisher.com terms and conditions…” and then click “Submit Order”.
You should now see a screen that says “Thank you for your order!” at the top. The “[email protected]” account will also have received a confirmation email (but you don’t need to check this or anything). You are FINALLY done. Log out of the account and go on with your day.