Rasmalo - Portugal
White Stork (Ciconia ciconia) nesting colony by the side of the road at Rasmalo in the Algarve.
Plus at least one in black plumage,
Lagos dos Salvados - Portugal
Ballast Toadflax (Linaria spartea)
Pink Pirouette (Silene colorata)
White Spanish Broom (Cytisus multiflorus)
Waxbill (Estrilda astrild)
Quinta Da Rocha - Portugal (3)
Corn Bunting (Emberiza calandra)
Snipe (Gallinago gallinago)
Common Sandpiper Actitis hypoleucus)
Spoonbill (Platalea leucorodia)
Chichchaff (phylloscopus colybita)
Cattle Egret (Bubulcus ibis)
Black-winged Stilt (Himantopus himantopus)
Phlomis purpurea
Cistus sp
Smilax aspera
Lichen Sp
Yellow Bee Orchid (Ophrys lutea)
Mirror Orchid (Ophrys speculum)
BumbleBee Orchid (Ophrys bombyliflora)
White Stork (Ciconoa ciconia)
Cape St Vincent - Portugal (3)
Lichen Sp?
Snail Sp?
Bee Sp?
Tick Sp?
Praia de Armoreira - Portugal
On the road down to the sea
Aljezur - Portugal
Lichen Sp
Pellia Sp
Fossombronia Sp
Fissidens Sp
Mediterranean Nettle (Urtica membranacea)
Green Winged Orchid (Orchis morio)
Lesser Snapdragon (Misopates orontium)
Cage Fungus (Clathrus ruber)
Ant Sp
Barbary Nut (Gynandris sisyrinchium / Moraea sisyrinchium)
Cley
Brent (Branta bernicla)
Lecanora sulphurea?
Pyrrosphora quercina
C+ orange (right)
KC+ orange (left)
UV+ orange
?
C+ Rose
Dirina massillienis f sorrediata
DNA Wheatfen
The Norfolk Fungus Study Group
DNA team
at work at
Wheatfen
under the watchful gaze of
Wheatfen's founder
Ted Ellis
Working surfaces are cleaned and sterilised with 10% thin domestic bleach in tap water with a drop of washing up liquid.
This is vital to prevent contamination, particularly if there of risk of amplified DNA on the surfaces.
A variety of pipettes, epindorf tubes & gloves needed for the process
Preparing numbered 1.5ml epindorf tubes with 100µl of extraction buffer ready for the samples.
Fungal specimens submitted in labelled packets
Using sterilised tweezers, razor blades and needles, and sterilising instruments in bleach between specimens,
small samples of fungal tissue are cut or torn off from the samples for testing.
Each sample is then placed in an appropriately numbered 1.5ml epindorf tube containing extraction buffer as mentioned above.
The samples are then ground with a sterile (bleached pestle) until the tissue is mostly pulverised.
Once ground teach tube has a further 400µl of extraction buffer added to it.
A seperate set of similarly numbered tubes have 1000µl of wash Buffer added to them.
Preparation of the 5x HOT FIREPol Master MIx
152µl of PCR Grade Water
40µl 5x HOT FIREPol Mix
4µl of prepared ITS1F primer at working concentration
4µl of prepared ITS4 primer at working concentration
Numbered tubes are then prepared, each containing 20µl of this solution
This is the moment when it all comes together and DNA is extracted.
1 A dipstick is dipped into the epindorf tube containing the sample with the extraction buffer three times.
2 The dipstick is then dipped into the tube containing the wash buffer three times
3 The dipstick is then dipped into the PCR mix tube three times to release the DNA
The PCR machine where the prepared samples complete with primers and FIREPol Mastermix are put through a thermal cycle
to amplify the DNA present.
Lab book recording
We've done it
Preparation for GEL electrophoresis
Ling Common
Bristly Haircap Moss (Polytrichum piliforum)
Hoary Fringe-moss (Racomitrium canescens)
Cladonia furcata 
Cladonia Sp?
Mossy Stonecrop (Crassula tillaea)
Peltigera Sp?
Heath Plait-moss (Hypnum jutlandicum)
The small white dots in this image are actually a tiny Lichen Coenogonium pinetti
Forked Veilwort (Metzgeria furcata)
Lichen Sp?
Ramalina farinacea?
Rustwort (Nowellia curvifolia)
Bristly Haircap Moss (Polytrichum piliferum) ♂︎plants
Chaenotheca ferruginea
Lateral Cryphaea (Cryphaea heteromalla)
Swan's neck Thyme Moss (Mnium hornum)
Cape Thread-moss (Orthodontium lineare)
Mueller's Pouchwort (Calypogeia muelleriana)
Shallow notches in the underleaves.
A lichen also present Diarthonis spadicea
Minute Pouncewort (Myriocoleopsis minutissima)
Creeping Fingerwort (Lepidozia reptans)
Lichen Sp?
Silesian Feather-moss (Herzogiella selgeri)
Arthonia atra
Lichen Sp?
Capsule without hairs from Ulotta crispa group
Frizzled Pincushion (Plenogemma phyllantha)
Ciliated Fringewort (Ptilidium ciliare)
Dilated Scalewort (Frullenia dilatata)
Two-horned Pincerwort (Cephalozia bicuspidata)
Knieff's Feather-moss (Leptodictyum riparium)
Forcipated Pincerwort (Cephalozia connivens)