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in this video I'm going to show you
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everything you need to know to get
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started with plant tissue culture so you
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can start cloning thousands of plants
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right from your home all of the supplies
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that I use for this tutorial will be
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linked in the description below so that
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you can easily replicate the process
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yourself the first thing that we're
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going to need is media for the plants to
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grow in tissue culture media is really
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important because it provides all of the
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necessary nutrients vitamins and
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hormones needed for the plant to grow
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outside of its natural environment the
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media which is is this clear substrate
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is typically made of five ingredients
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the first ingredient is distilled water
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which makes up the bulk of the media the
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second is migi and scog a basil salt
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medium which is usually abbreviated to
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Ms the third is sugar this gives the
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plants the energy that they need to grow
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fourth is a jelling agent to turn the
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media from a liquid to a semi-solid this
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will usually be either AAR or jeling gum
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and fifth is plant growth Regulators or
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pgrs which are plant hormones if you
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live in the United States all of these
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ingredients are really easy to get if
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you live elsewhere I would recommend
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joining the plants and jars Discord
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server we have people doing TC all over
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the world who can help use Source
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supplies no matter where you live the
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quantities of these ingredients to make
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one liter of tissue culture media will
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differ depending on the protocol that
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you're following a protocol is simply a
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set of instructions for culturing a
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particular type of of plant and it'll
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provide the exact recipe for preparing
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the media you can find protocols on
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websites like research gate or Google
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Scholar and I share and test out a lot
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of different protocols here on the
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channel here's an example of a tropical
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house plant media recipe that I use all
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the time this media works great for
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filad dendrin monsteras Epi pums I
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haven't tried it for singon or for
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alloces yet but I do suspect it would
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work for those as well following these
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instructions I add 800 ml of distilled
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water to my container then I add 4.54 G
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of Ms 30 g of sugar 3 G of jelling gum
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and then my plant growth Regulators
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which are 1 mg per liter of Bap and 0.1
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mg per liter of IBA keep in mind plant
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growth Regulators are technically
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pesticides I would always recommend
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reading safety data sheets for whatever
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chemicals you're working with to
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familiarize yourself with how to handle
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them properly store them and also how to
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dispose of them once everything is added
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top off the water to the 1 lit Mark I
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mix everything together on a magnetic
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stirer the one that I'm showing in these
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clips is pretty expensive but I'm going
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to link a more economical option below
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that I've used and that I would
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recommend the pH for tissue culture
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media is super important most protocols
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require the pH of the media to be
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between 5.6 and 5.8 I adjust the pH
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using hydroponic up and down until it's
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at 5.7 this is not the most scientific
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way to do this but it's easy and it
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works then you're going to pour the
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media into individual containers the
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level that you fill them should be about
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the width of your index finger and the
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containers need to be either glass or
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polypropylene and any Lids need to be
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made of polypropylene or else they going
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to melt during the next step of the
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process which is Media sterilization so
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the media needs to be completely sterile
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for TC if there's even one little
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surviving cell of bacteria or one Spore
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of mold your culture will get
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contaminated and you'll have to start
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over it's very sad when that happens
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we're going to pressure cook the tissue
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culture media for 15 minutes at 15 psi
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in an old school conventional style
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pressure cooker make sure not to secure
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the lids of the containers before
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running the pressure cooker just run
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them with the lids resting slightly on
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top and then once you open the pressure
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cooker you just want to snap the lids on
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immediately we're also going to need to
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sterilize a set of forceps I buy these
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sterilization
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pouches these are intended for dentist
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offices I'm pretty sure but they work
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great for tissue culture the forceps
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also get sterilized for 15 to 20 minutes
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at 15 psi so the same exact way that you
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sterilize the media most protocols will
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let you know what part of the plant to
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use for TC and give you instructions on
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how to sterilize that part of the plant
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usually with bleach 99% of the time this
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is where the contamination happens if
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you're new to tissue culture I would
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suggest instead of starting with an
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existing plant start with one that's
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already in tissue culture this ensures
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that the plant is already sterile from
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the beginning of the process and it
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allows you to just put it into
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multiplication media to get it to start
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cloning itself again in the past few
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years we've seen a big increase in the
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amount of people companies and
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Laboratories selling plants that are
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still in tissue culture media on the
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internet the philad Dundon red Anderson
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that I'm using today for the
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demonstration is from a company called
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me Greenery export they were kind enough
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to send me the plants for the video this
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is the owner her name is Fern they also
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sent me a few other plants as well which
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was really kind of them in terms of
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other places to Source TC plants I know
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EAA has a tissue culture catalog if you
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ask them for it aoid Asia also sells
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plants in TC Orange Lake Nursery in
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Florida doesn't do TC but they do
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contract with the TC lab to sell plants
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and then there are tons of aquatic
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plants available in tissue culture these
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are some anubias that I got from
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aquarium plants Factory the Nuance when
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you buy tissue culture plants for this
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process is that you really want them to
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be in a container or a glass jar like
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this you don't want them to be in a
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baggie like this the baggie is much
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clumsier to work with with and you'll
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have higher instances of contamination
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for anyone interested in pursuing TC as
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a long-term hobby I would recommend
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eventually picking up a lemoner Flo Hood
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like the one you see behind me it just
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helps keep everything sterile and you'll
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have a lot less contamination than using
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a still airbox but a still airbox is
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what I will be using for this
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demonstration and you can still have
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success with it if you're very careful
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the more you get into tissue culture the
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more your studio apartment starts to
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look like a evil sciencetist layer it's
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inevitable so don't try to fight it to
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get started with my transfer I spray
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everything down with 70% alcohol
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including my own arms and gloves inside
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of the still airbox I have my containers
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of fresh multiplication media that we
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made earlier the plants from me Greenery
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export my forceps and a pair of scissors
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which had both been autoclaved in the
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pressure cooker before starting and also
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a cheap glass Beed sterilizer the glass
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Beed sterilizer is where I store my
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forceps when I need to put them down it
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is technically meant to be used by nail
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technicians but today I'm using it for
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the pursuit of science once everything
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is set up the transfer process is really
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quick I essentially just take the filad
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dendrin out of the container it came in
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and transfer it into new fresh
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multiplication media you want to work
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quickly avoid leaving containers open
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unnecessarily and never put down the lid
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of a fresh media container just hold it
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in your non-dominant hand another tip is
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to avoid Reaching Your Hand hands and
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arms over the top of any open TC
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containers to prevent contamination as
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much as possible I repeated the same
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process with the monstera but it's
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really hard working with the bag the
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plant did touch the outside of the bag
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as I've removed it with my forceps and
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unfortunately became contaminated so
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again I really don't recommend using the
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bagged plants for TC make sure to wrap
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the containers with plastic wrap so they
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don't get contaminated instead of buying
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plastic wrap that pre-cut I just cut it
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into 1 to 2 in sections with a PVC pipe
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cutter to save money and that's what I
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Ed to seal the containers so it's been
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about 3 weeks since I filmed that demo
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as I mentioned the monstera did get
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contaminated but the fil adenon red
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Anderson has absolutely no contamination
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you can see how clear that media is I
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don't see any shoots growing from it yet
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but we should have some shoots hopefully
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forming very soon once a shoot forms
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that is where you'll start to see kind
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of baby plants start to come out of it's
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hard to explain I'll insert some photos
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so you can tell what I'm talking about
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but you'll need to subculture your
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plants every four to 6 weeks so that
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just means moving them from this media
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to New Media even if it's the same
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multiplication media recipe you still
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need to keep moving the plants otherwise
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you'll risk the containers filling up
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with ethylene gas and killing the plants
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once you have enough plants and you're
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satisfied you would either put them into
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rooting media or you could just take
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them out of tissue culture and acclimate
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them to growing in soil if they're large
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enough and that's it I keep my plants
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under grow lights for 16 hours a day on
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a timer this process I know seems really
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intimidating at first um I made a lot of
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mistakes when I was first getting
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started 2 years ago like a lot of
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mistakes but eventually once you start
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to get the hang of TC it becomes the
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coolest hobby ever and it is so
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worthwhile if you want to learn more
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about tissue culture I have a lot more
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tutorials on my channel that go much
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more in depth than this one I'd also
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recommend reading the book plants from
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test tubes it's a five out of five
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absolutely fire and finally I'd like to
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thank the sponsor of today's video plant
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cell technology plant cell technology is
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basically a One-Stop shop for tissue
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culture supplies they sell everything
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you need to make media like we did in
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this video today they also have a tissue
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culture masterclass in case you want to
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jump start your tissue culture learning
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experience I've taken the in-person one
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and it was genuinely a really awesome
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experience and I learned a lot I would
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definitely recommend it the next
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inperson class is going to be in
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Washington DC from June 7th to June 9th
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and if you're unable to travel there in
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person they also do an online class and
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the next online class will be July 27th
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and July 28th I do have a code for both
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those classes if you're interested it is
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plants and jars and it'll give you 10%
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off of the price mind blank no thoughts
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bye guys guys
