Participants involved in the Fish-T1K Project are expected to provide tissues or RNA samples for target fish species. The following protocols forcollecting, sampling and transportation are recommended:
All tissues should be documented with a voucher specimen. Preserved specimen is preferred, but digital photo voucher is also acceptable.
Record as much information as possible for each specimen (date of collection, location, size, sex, etc.) in the metadata sheet.
Vouchering procedures. After tissues are collected, voucher specimen should be fixed in a buffered 10% formalin solution (or 3.7% formaldehyde solution) for 2-10 days and then transferred to 70% ethanol for long-term storage. Large specimens should have formaldehyde injected into body cavities to ensure uniform fixation of muscle, gut cavity and brain tissue.
Please provide 10 μg of genomic DNA or ~ 0.5 g of fin or muscle tissue for each target species, or more if possible. Tissues collected need to be preserved in ethanol or frozen in liquid nitrogen and subsequently stored at -80°C.
Tissues from 4 organs (gill, eye, muscle and liver) are required. All tissues should be preferably collected from a single individual. RNA extracted from different tissues of one species will be used to construct libraries for RNA-seq.
Tissues may be preserved in RNAlater and stored at room temperature, or frozen in liquid nitrogen and stored at -80°C. Initial flash-freezing in liquid nitrogen is strongly recommended for new acquisitions when field conditions permit. For collection in RNAlater, larger tissues should be cut into small pieces (~5 mm) to permit adequate fluid penetration. Use 1:5 ratio of RNAlater (e.g., 0.5g tissue: 2.5 ml RNAlater).
Please provide 10 μg of total RNA (OD260/280 ≥ 1.8, OD260/230 ≥ 1.8; 28S/18S ≥ 1.0, RIN ≥ 7.0) or 1~2 g of tissue for each target species, or more if possible. For smaller size fishes, tissues less than 1 g are also accepted provide they can yield ~10mg total RNA.
Transport tissues to the lab in liquid nitrogen or dry ice. Frozen tissues should be stored in -80°C freezer or in liquid nitrogen.
All tissues used in the project must conform to the laws and statutes relating to the collection and manipulation of biological material in the respective countries. In particular, proper permits should be obtained for collecting and transporting rare and endangered species.
To send the tissue samples to BGI-Hong Kong (the samples must be non-infectious), an import license will be required. Please help to prepare the following information and send to firstname.lastname@example.org for application:
Frozen tissues should be transported in adequate quantity of dry ice that will last until the delivery of the sample to Hong Kong. Transport permits are required for dry ice, which is classified as "Restricted Articles". Confirm updated regulations prior to shipping.
Since it normally takes 3-4 days to ship from US/Europe to HK, make sure you store the samples in a tightly-sealed styrofoam box, with plenty of dry ice (at least 15Kg) to guarantee that some dry ice will be in the box when the sample arrives to HK.
RNAs are highly recommended to be stabilized in Gentegra or similar regents to be transported at room temperature because this will be the most convenient and cost-effective way. Unlike tissues, RNAs do not need import permit. In addition, ambient temperature shipping uses no dry ice (which is regarded as dangerous goods in air traffic so that particular shipping document as well as handling charge will be needed).
If RNAs are isolated from endangered fish species and restricted under CITES, CITES permit will be needed for exportation/ importation.
Please prepare an excel file with detail information of your RNA samples. It must contain the following information:
While everything is settled, please ship the precious samples to the following address in the table below. All samples coming overseas are firstly sent to BGI-Hong Kong Sample Management Center and then transfer to BGI-Shenzhen.
|Consignee's name||Attend to: Ms. April Wong. BGI Tech Solutions (Hong Kong) Co., Ltd|
|Consignee’s address||2/F, 16th Dai Fu Street, Tai Po Industrial Estate, Tai Po, Hong Kong|
|Contact number||852-3610 3511; 852-9683 9836|
|Notes||Please be informed that HK has no postal/zip code. To avoid delay, please advise the client not to put China/CN in the address. In case "Country" must be filled, please just put down "Hong Kong".|
Please ship the samples on the first 1-2 days of the week, such as Monday/Tuesday, in order to avoid shipping time being extended during weekend. After sending out the shipment, please provide us with the name of the express and the tracking no.
As soon as we receive the shipment, we’ll write back to you to inform its safety and begin to extract/recover RNA samples and have them tested (if there are any special instructions you need us to follow, please point them out in your mail). Usually, the quality assessment report of the samples will come out and be sent to your e-mailbox within two weeks (sometimes longer because of holidays) and we can discuss further sequencing plans.