If you want to set up a lab for bird sexing, what equipment do you need to get started? And what should you consider when starting out?
This article covers the essential equipment, reagents, and plastic consumables that you’ll need to set up a bird sexing workflow for the first time, and also what you’ll need to restock your supplies when you’re running low on reagents.
What essentials can you buy from us?
- A Bento Lab or equivalent lab equipment. We have two Bento Lab models, Pro and Entry (see here for a detailed comparison):
- Bento Lab Pro: more versatile, can use touchdown PCR, with an adjustable high speed centrifuge ( (8,000 ×G), and adjustable gel voltage
- Bento Lab Entry: can’t do touchdown PCR, has a fixed speed centrifuge ( (2,700 × G), and fixed gel voltage
- Our Bird Sexing Bundle (for 48 birds)
- A top-up of enough DNA extraction reagents, PCR reagents, and plastic consumables for a given number of samples (e.g. 100 – 400) (see below for details)
What essentials do you source elsewhere?
- A microwave oven for melting agarose gels
- A freezer for storing reagents
- 1 L of distilled/deionised water for making 0.5× TBE electrophoresis buffer from the 10× TBE stock solution
- An empty 1 L bottle to make up 0.5× TBE electrophoresis buffer from the 10× TBE stock solution
- Permanent markers for labelling tubes
- Nitrile/latex gloves
- A sharps bin or sturdy sealable container
- A sealable plastic container for plastic waste/tip bin
- A small bin/bin bag
- A decontamination solution: one part thin domestic bleach and nine parts tap water, with a drop of detergent added, ideally in a spray bottle for easier use
- Kitchen roll or paper towels for spills and drying purposes
What are the nice-to-have’s?
Buy from us
- Add-A-Lane comb to allow you to run up to 24 lanes in a gel
- An extra 20 µL micropipette for working only with PCR products, to minimise PCR product contamination risk
Buy elsewhere
- One or more large plastic trays to work on
- Kitchen roll/paper towels/aluminium foil to work on
- Small plastic bags to store DNA extracts and PCR products
- Small boxes to store reagents in the freezer
- Larger plastic boxes for equipment (e.g. 9 L “Really Useful Boxes”)
- A small electronic kitchen weighing scale (accurate to 1 g or less) for measuring electrophoresis buffer
- Tweezers (e.g. ×10)
- Double-edged razor blades as a cheap disposable tool for cutting feathers
What are the essential reagents and consumables?
The following reagents and consumables are all used in the bird sexing workflow, and can be found in the Bird Sexing Bundle in quantities sufficient for sexing 48 birds (with some reagents left afterwards).
Once you have used them up you can buy them as individual items from our store.
| DNA Amplification | Description | Amount used per PCR |
| 5x HOT FIREPol Master Mix | 5× concentrate of the PCR reaction mix, lacking bird sexing primers, water, and the template DNA. | 4 µL per 20 µL PCR |
| PCR grade water | Extremely pure water with no contaminants | 12 µL per 20 µL PCR |
| Bird sexing primers | DNA “tags” defining the region to be amplified. Read here how to decide which primers are suitable for your bird species. | 2 µL per 20 µL PCR |
| DNA Extraction | Description | Amount used per extraction |
| HotSHOT DNA Extraction Kit | An inexpensive, easy to use DNA extraction method. Read how to use it for feathers or blood spots here. | 75 µL of extraction solution and 75 µL neutralising buffer |
| Agarose gel electrophoresis | Description | Amount used per gel (8 samples or 18 samples with the add-a-lane comb) |
| 10x TBE buffer (diluted to 0.5× before use) | A buffered solution used for casting and running agarose gels. Bought as a 10× stock solution, it is diluted ×20 to make a 0.5× TBE buffer. | 0.5× TBE: 25-50 mL per gel, and 40-50 mL running buffer |
| Agarose tablets | A concentrated agarose tablet that is dissolved in 0.5x TBE buffer to form an agarose gel | 1× 0.5g tablet per 25 mL gel, or 2× 0.5 g tablets per 50 mL gel |
| GelGreen DNA stain (or other blue-light compatible stain) | A dye that fluoresces green when bound to DNA and exposed to blue light | 2.5 µL per 25 mL gel, or 5 µL per 50 mL gel |
| 100 bp DNA ladder | A mixture of different-sized DNA fragments in 100 bp increments, used to determine the size of your DNA fragments | 5 µL per lane (use two per gel row at either end, or one in the middle of the gel) |
| Plastics | Description | Amount used per batch of 8 samples plus 2 controls |
| PCR tubes | 0.2 mL tubes used for HotSHOT DNA Extractions and PCR | 20-30 PCR tubes |
| 1.5 ml tubes | 1.5 mL tubes for making batch PCR mixes | One tube per batch PCR mix |
| 2-200 uL pipette tips | Pipette tips used for all pipetting steps with both pipettes | ~28 tips without a DNA extraction dilution step, and ~38 tips with a dilution step. |
Topping up the Bird Sexing Bundle for larger numbers of samples
If you want to sex more than 48 birds, then you can top up the bird sexing bundle to a larger number of birds by buying additional items, for example for 100 birds, 200 birds, and 400 birds as detailed in the table below.
Please note that these calculations are based on running eight samples plus two controls plus two DNA ladder lanes in a single batch, i.e. a single row gel. If you are using the Add-a-Lane comb, you can run more samples per gel and therefore use fewer reagents for agarose gel electrophoresis per sample.
| Item | 100 birds | 200 birds | 400 birds |
| HotSHOT DNA Extraction Kit | NA | NA | NA |
| 5x HOT FIREPol Master Mix | 1 × 400 µL tube | 1 × 1000 µL tube | 2 × 1000 µL tube |
| PCR grade water | 1 × 5 mL bottle | 1 × 5 mL bottle | 3 × 5 mL bottle |
| Bird sexing primers | NA | 1 × 400 µL tube | 2 × 400 µL tube |
| 10x TBE buffer | 1 × 50 mL tube | 2 × 50 mL tube | 4 × 50 mL tube |
| Agarose tablets | 1 × 10 tablets | 3 × 10 tablets | 8 × 10 tablets |
| GelGreen DNA stain | NA | 1 × 100 µL tube | 1 × 200 µL tube |
| 100 bp DNA ladder | NA | 1 × 200 µL tube | 2 × 200 µL tube |
| PCR tubes | 2 × 100 tubes | 1 × 500 tubes | 1 × 1000 tubes, and 1 x 100 tubes |
| 1.5 ml tubes | NA | NA | NA |
| 2-200 uL pipette tips | 2 × boxes of 96 tips | 7 × boxes of 96 tips | 17 × boxes of 96 tips |
Buying reagents and consumables without the Bird Sexing Bundle
If you want to buy reagents and consumables without the Bird Sexing Bundle, for example if you are restocking reagents or already have pipettes and the other equipment in the Bird Sexing Bundle, you can use the table below to work out what you need for a given number of samples.
Please note that these calculations are based on running eight samples plus two controls plus two DNA ladder lanes in a single batch, i.e. a single row gel. If you are using the Add-a-Lane comb, you can run more samples per gel and therefore use fewer reagents for agarose gel electrophoresis per sample.
| Item | 100 birds | 200 birds | 400 birds |
| HotSHOT DNA Extraction Kit | 1 × HotSHOT DNA Extraction Kit | 1 × HotSHOT DNA Extraction Kit | 1 × HotSHOT DNA Extraction Kit |
| 5x HOT FIREPol Master Mix | 1 × 1000 µL tube | 1 x 1000 µL tube, and 2 x 100 µL tubes | 2 × 1000 µL tubes, and 1 × 400 µL tubes |
| PCR grade water | 1 × 5 mL bottle | 2 × 5 mL bottle | 3 × 5 mL bottle |
| Bird sexing primers | 1 × 400 µL tube | 2 × 400 µL tube | 3 × 400 µL tube |
| 10x TBE buffer | 2 × 50 mL tube | 3 × 50 mL tube | 5 × 50 mL tube |
| Agarose tablets | 3 × 10 tablets | 5 × 10 tablets | 10 × 10 tablets |
| GelGreen DNA stain | 1 × 100 µL tube | 1 × 200 µL | 1 × 200 µL, and 1 x 100 µL |
| 100 bp DNA ladder | 1 x 200 µL tube | 1 x 400 µL tubes | 1 x 400 µL tubes |
| PCR tubes | 1 x 500 tubes | 1 x 1000 tubes | 1 x 1000 tubes, and 1x 500 tubes |
| 1.5 ml tubes | 1 x 100 tubes | 1 x 100 tubes | 1 x 100 tubes |
| 2-200 uL pipette tips | 5 × boxes of 96 tips | 10 × boxes of 96 tips | 20 × boxes of 96 tips |
Setting up a lab workspace
When setting up a lab workspace, consider the following and whether you need to buy anything to make a satisfactory working environment:
- Availability of plug sockets (you may need an extension cable)
- Good lighting (you may need to buy a lamp to improve visibility)
- A decontamination-compatible surface (you may need to buy a plastic table, or plastic table cloth, or trays to work on).
- Proximity to a microwave oven for melting agarose gels (you may need to buy a microwave oven or a rice cooker)
- Where you will conduct each step of the workflow (you may like to buy a separate table for sampling specimens).
Please note that you don’t necessarily have to do all steps of the workflow (sampling, PCR set-up, running the PCR, gel casting, gel running and visualisation) in the same place.
In fact, to minimise the risk of PCR product contamination, it can be very useful to do pre-PCR steps (sampling and set up the PCRs) in a separate area or on a separate tray to where you are doing the PCR and post-PCR steps (casting, loading, running, and visualising the gel).
Read our article on setting up a lab workspace here.
Storing equipment and reagents
You may also want to buy some storage solutions for your equipment, reagents, and consumables.
Cold storage
A small number of items are best stored at -20 °C in a freezer to maximise their working life. These are:
Pre-PCR
- PCR master mixes (5× HOT FIREPol Master Mix, 5× FIREPol Master Mix)
- Primer mixes
- PCR grade water (not essential, but freezing will minimise the risk of microbial growth)
Post-PCR
- DNA ladder (not essential, but it might extend the reagent shelf-life)
- DNA extractions (if you are keeping them)
- PCR products (if you are keeping them
Store cold storage products in a non-frost-free freezer to maximise their working life. If you store in a frost-free freezer, package reagents in a protective box/packet to minimise freeze-thaw during defrost cycles.
Keep post-PCR items (DNA ladder, DNA extractions, PCR products) separate from pre-PCR items (everything else) to reduce the risk of contamination by PCR products.
Ambient storage
Store in plastic boxes
- HotShot DNA Extraction Kit
- 10× TBE buffer (and 0.5× TBE buffer)
- Distilled water
- Agarose tablets
- GelGreen DNA stain
- Micropipettes
- Pipette tips
- 0.2 mL PCR tubes
- 1.5. mL tubes
- Miscellaneous equipment (waste bins, kitchen roll, tray, etc.)
If you have any questions, please get in touch!
Please let us know what other resources, advice, and tips and tricks for using Bento Lab that you would like us to produce in the future!
Looking for advice on using Bento Lab?
Book a free consultation or ask a question.