HomeGadgetsA Guide to Properly Filling 96-Well Plates

A Guide to Properly Filling 96-Well Plates

Learning your equipment is one of the top tips for success in the chem lab, and 96-well plates are no different.

Imagine your big day has finally arrived: your first independent experiment as a junior researcher in the lab. With steady hands, you fill tiny droplets in each of the ninety-six wells on a plastic plate. Finally, well 96 is filled, or so you thought.

As you withdraw the pipette from the last well, your heart sinks. Your plate is a picture of inconsistency: a few wells brimming, some barely half-full, and some overflowing, mixing into their neighbors. The realization hits you like a bus: the way you filled your plates could invalidate all of your data.

To make sure your PCR tests yield the most accurate results possible, read this guide on how to handle 96-well plates-no more guesswork and no more mistakes.


Sure, maintaining sterility is paramount in any chemistry lab, but this is especially true when filling 96-well plates.

If your sterile practices aren’t enough, contamination will rear its ugly head. You know, the pesky, unwanted guest that messes up your lab results-the science equivalent of that person crashing your party and eating all the guacamole.

To keep those party-pooping contaminants from turning your experiment into a disaster zone, store your deep well plates in dry dry conditions.

If a protocol calls for plate surface treatment (such as gelatin coating), this process should happen in a disinfected area. The treated plates can then be stored in a culture hood.

All equipment coming into contact with the 96-well plates, such as pipettes, should be purified before use. And, before handling the plates, gloves should be changed or cleaned with seventy percent ethanol.

Preparing 96-Well Plates

Different experiments require different preparations. For example, if you’re using the plates for adherent cell cultures, the first step is coating the wells with the required molecules-usually proteins. If you skip this step, the cells won’t attach.

Each well requires a specific amount of coating solution. Some products say 0.1 mL is more than enough. Even so, pull out your magnifying glasses and verify whether this is the case for your product.

Filling the Wells

This part requires deft laboratory skills. Your movements must be so precise that you make Bill Nye look like a preschooler playing with a DIY volcano kit.

Using a micropipette, add the required amount of solution to the well plates.

It’s important to fill along the same line. If you start filling in the top left corner, ensure the subsequent wells are filled on the same side. This habit results in more consistent data and will make it easier to read the plate later on.

Well Plate Sealing

Proper sealing prevents differential evaporation of samples in the thermocycler. If not controlled, improper vaporization could alter the pH of the solution and impact enzymatic functions. This would disrupt the results.

Plate sealing films are transparent heat-sealing covers generally used for 96-well plates. These are most suitable for PCR reactions and storage.

Aluminum sealing films give a more robust seal. Most people heat-seal them to the plate. As a good barrier against evaporation, they’re able to maintain the internal environment of the well during storage or reaction processes.

Soft-cap mats are a more economical choice for short-term storage, such as when setting up a PCR test.

Avoiding Common Mistakes

First up, we’ve got cross-contamination. This occurs when a solution spills over into other wells from a pipette.

To cut the risk of cross-contamination, treat your pipettes like fresh gossip; don’t let them spread just anywhere. Real winners change pipettes as soon as they start using a new solution.


Staying consistent while filling over ninety wells is a nightmare; even the slightest disruption can leave you frustrated. Not only can the lack of consistency transform your cheerful lab hours into a reel of nightmares, but it also harms the reliability of your results.

Imagine being part of a choral performance. Your group is belting out Beethoven’s ninth symphony, but the tenor, in a bold plot twist, has decided to freestyle. The harmony is disrupted, the performance is thrown off-balance, and the audience is perplexed.

That’s precisely what a lack of consistency can do to your scientific work.

Consistency is the hallmark of truth in the laboratory. So, whether it be your pipetting technique, the exact measurement of reagents, or the precise control of variables, strive for consistency.

Types of Well Plates

The sheer number of well plates available online can overwhelm you, making it easy to choose the first option that comes along. But remember, one size does not fit all.

Consider the volume requirements of your experiment. When it comes to large volumes, use deep well plates.


Ensuring accuracy during laboratory tests is a tricky endeavor for sure, especially during repetitive tasks like pipetting. Frustration can easily bubble up and cause your hands to shake.

So, how can you cultivate patience in the lab while keeping annoyance under the calibrated lid?

First, be present and focus on what you’re doing. No, we’re not suggesting that you make the lab your meditation grounds, but being mindful of each step helps reduce errors and, subsequently, frustration.

Remember to take things slow. Cultivating patience often involves pacing yourself. With each steady action, you’ll build resilience against frustration.

Also, have some free time in your schedule. This means not packing your timetable so tightly that one slip-up sends your whole day spiraling into an abyss of rushed inaccuracies. Taking breaks reduces pressure and gives you the freedom to be less anxious.

Achieving Precision and Success in the Lab

Properly filling 96-well plates requires patience and, more importantly, laboratory skills. This art could be the difference between a sound experiment and one that’s misleading. So, always practice sterile techniques to avoid contamination and ensure consistency when filling.

If you want to maximize your test scores, it’s important to have the right equipment and lab tools. Explore our gadget sections for more devices that would be useful in your lab.

Daniel Robert
Daniel Robert
Daniel Robert is a multi-talented author at thetechdiary.com, particularly interested in business, marketing, gaming, entertainment, technology and more. His diverse background and love for learning have allowed him to write on various topics. With a unique ability to craft engaging and informative content, Daniel has become a well-respected voice in online publishing.


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