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Tissue Culture

* Good Quality Phalaenopsis Flask Plants
In order to produce good quality Phalaenopsis flask plants, the following procedures are strictly and carefully performed for quality control and assurance.
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Strict Mother Plant Selection
Selecting mother plants for tissue culture propagation, the plants are being strictly selected by their flower, leaf, and overall plant appearances for good characteristics and without mutation.  Furthermore, the plants are tested by ELISA and RT-PCR for virus free material.  The mother plants are renewed with new plants regularly for the material vitality.
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Limited Sub-Culture Cycles
When the Phalaenopsis chromosomes are experiencing fast and multiple times of dividing, incomplete division can easily occur and result in mutation.  Limiting sub-culture cycles and coop with stable culture medium can prevent mutation.  Each meristem material is limited to 10 sub-culture cycles.
For Contract based tissue culture production, we are able to provide a production schedule according to customers’ request 6 months after receiving the spikes and numbers of sub-culture cycles are also determined to ensure production quality and quantity.
 
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Healthy Plant Production
Every spike of mother plants is tested by virus detection individually.  During the first stage (initiation of explants, establishment of mother plant from meristem of nodes in spikes), two times of RT-PCR virus testing are perform for ORSV and CymMV virus detection.  After the material is determined clean and non-virus, it can be used in tissue culture propagation.  The virus detection will be implemented again at the last stage (stage before acclimation) of tissue culture propagation for guaranteed delivery of virus free healthy flasks.  For any specific diseases or virus detection, please request prior to spike delivery.
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Barcode System
To avoid confusion of the varieties, barcode system has been carried out during tissue culture production.  From the reading of the barcodes during the operation process, the operator can ensure the labels on first stage bottles match with last stage bottles.
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Protocol of Spike Preparation, Packing and Shipping Procedures for Tissue Culture Propagation
There is a protocol for spike preparation, packing and shipping procedures for tissue culture propagation.  (Please click here for web link and file downloading.)   Please post the spike to our Tissue Culture Department in Taiwan within two days to obtain the best vitality of the meristem cells for good tissue culture material.
* Recommendations on Shipping and Deflasking
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Adjustment of Production and Shipping Schedule
If the production quantity or shipping schedule needs to be adjust, please inform us six month in advance for rearrangement of the production schedule.
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Glass Bottle or Plactic Bottle
If there is a preference for glass bottle or plastic bottle, please inform us three month in advance before the shipping date.  If no request is received, we will use most current available bottles.
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Acclimation Environment
The recommend acclimation environment is with light intensity between 70 to90μmol-1 and temperature range from 22C to 32C.  If you have a high light and high temperature acclimation environment, please deflask immediately to avoid flask plants from aging which may affect propagation quality.  In order to avoid excessive growth, please do not place the flask plants in low light and high temperature environment.
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De-flask Timing
Please de-flask within 10-14 days after receiving.  The flask plants are adjusted to the new environment while aging is avoided.  If you do not need us to perform acclimation for you, please also inform us two months the latest in advance before shipping date.
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Flask Plants Cultivation Techniques
For flask plants cultivation techniques, please refer to “Cultivation Guide of Phalaenopsis Plants.”
* Phalaenopsis Tissue Culture
Spike Preparation Standards, Packing and Shipping Procedures
NO.
 Steps Description
1
Preparation
  1. Verify the obtained variety and spike quantity.
  2. Take photos of flowers of each variety for reference.
  3. No mutation on any petals (discard any spike with mutated petals.)
2
Spike Removal from the Plant Body
1. Sterilize the blade with flam every time before removing a spike from the plant body.   2. Remove the spike from the plant body at the most bottom point.
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3. Categorize and mark the spike with the variety.
  4. Remove a little piece of spike off at the tip and rare end.  Seal with wax to eliminate dehydration.
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3
Packing
1. Save the first or second full bloomed flower and remove the rest buds off.
 
2. Cover the spike completely with newspapers while leave the flowers uncovered.  
3. Please remove the branches from the spikes and cover in newspaper as well.  Please make sure all the spikes are kept inside the newspaper wrap.
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4. After wrapping the spikes, please place the spikes in boxes with Styrofoam peanuts or newspaper scraps.  
5. Seal the box with tape carefully.
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4
Delivery
  1. Please mark the box with our company name and address and yours as well on the box.
    Royal Base Corporation
    No.88-1, Cingguang Rd., Wurih Township, Taichung County 41462, Taiwan.
  2. Please mark fragile and handle with care on the box.
  3. Please deliver through a professional forwarder.
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Note
  1. Stop watering or fertilizing one week before removal of the spikes.
  2. In order to keep freshness and vitality of the spike, please harvest the spike from the plant once the first bloom has fully opened.
  3. To eliminate virus infection between the spikes, please isolate the spikes from each other.
  4. Please place the spikes with open blooms on the top of the box to eliminate bruises on the petals.
  5. 【IMPORTANT】The spikes need to be kept between 15 ~ 20 C (56 ~ 68 F) to maintain freshness. Please inform your forwarder or freight carrier about the temperature requirement.
  6. Please inform us immediately after the spikes are shipped.  Please provide us the tracking numbers of the parcel if possible.
  7. Photos of the spikes will be taken after receipt and a copy will be sent to you as well for your reference.
 

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