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 1. What is DEG? |
DEG (Differentially Expressed Gene) is a gene that shows differences in mRNA expression
level between two or more RNA samples treated in different ways. |
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| 2. Where can DEG service be applied? |
Various researches such
as
Research on the disease related gene and diagnostic
marker
Research on the tumor marker gene and diagnostic
marker
Research on the gene reacting to specific
drug/hormone treatment
Research on the gene related to development
and differentiation
Research on the gene related to aging or metastatic
cancer
Research on the candidates of anti-cancer
agent or new drug
Research on the various biomarkers
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3.
What are the previous competitive methods
of DEG identification and
what is the advantage
of GenfishingTM DEG analysis? |
The previous DEG identification methods can be categorized by two groups, PCR based methods and
hybridization based methods
PCR based methods: differential display (DD-PCR), serial analysis of gene expression (SAGE)
Hybridization based methods: differential
hybridization, subtractive hybridization,
microarray (DNA chip) DD-PCR or DNA
chips were the most popular methods to identify
DEG, but problems such as false positive
products and low reproducibility have been
reported. Seegene developed a new DEG identification
method using GenefishingTM technology which
solved the problems of false positive and
low reproducibility.
In addition, the GenefishingTM DEG discovery
method is more economical and easier than
previous DEG
identification methods since DEG can be detected
on the agarose gel. |
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| 4. What gene coverage does Genefishing¡Ëa service have? |
For the GeneFishing¡Ëa DEG service, average gene numbers amplified by one primer pair are 10 to 15.
Therefore, 1.200 to 1,800 genes (100bp ~ 2.0 kb size) can be amplified by 120 primer pairs of full screening
service. This is relatively lower gene coverage than other methods such as DNA chip or SAGE. However,
the Genefishing¡Ëa DEG method has a strong advantage of no false products, discover of novel gene, detection
of rare transcripts, less expensive, and a simple procedure in comparison to the other previous methods |
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| 5. What criteria was used to select DEGs from the result report of GeneFishing¡Ëa service? |
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For the most accurate data, our reserchers repeat the experiment and identify DEGs confirmed by a second experiment with their expertise. |
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| 6. Where can I get detailed information? |
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Please visit the Seegene¢®?s webpage (www.seegene.com) or contact Seegene by telephone (301-990-4931) or fax (301-990-4932). |
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| 1.What criteria was used to select 10mer core region of arbitrary primer? |
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Possible combinations of 10mer core region are 10 multiplication of 4 (A, T, G, C), which is more
than a million. Among the combinations, ideal combinations up to 120 pimers (full screening) were
selected based on the amplification size and frequencies occured in most eukaryotes. |
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| 2. Can I get the primer information? |
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Among the 120 arbitrary primers, we can only provide information from the first to 20th arbitrary primer.
(arbitrary ACP1 to arbitrary ACP20) |
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3.
Is there any primer pair that can detect specific
genes
better among Genefishing¡Ëa
arbitrary primers? |
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Since arbitrary primers are randomly designed, there is no primer pair that detects specific genes better. |
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| About the free-trial screening service |
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| 1. What service can I expect in regard to the free-trial screening? |
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Among the total 120 primers, 20 primers are selected randomly and used for the free-trial screening.
The free-trial screening result (gel photo) and recommendation file for the next experiment will be
provided to clients. (However, PCR product will not be provided) |
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| 2. How many DEGs are identified with the pre-trial screening service? |
There is no rule. However, if morphology and phenotype shows big differences,
there is a greater possiblity to produce many DEGs. |
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| 1. How many samples can I request for free-trial screening service? |
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Only two samples can be accepted for the free-trial screeening and we will charge for additional samples.
Among the two samples, one should be the control to be compared. |
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| 2. How can I prepare the samples? |
We recommend RNA samples for the DEG service.
However, our laboratory also offers RNA preparation
service (Cat. No, S1001) and you can also
send tissue or cell type samples. In these
cases, infectious
materials can not be accepted under any circumstances.
RNA sample: Require 50¡×¢Ò total RNA (con. 0.5
to 1.5¡×¢Ò/¡×¢®) in DEPC water. Please send your
RNA samples with RNA quality confirm data
(RNA extraction procesure, formaldehyde gel
photo, OD value) because the result of DEG
service will be greatly influenced by the
quality of RNA.
Tissue sample: Immediately freeze (-70¢®¨¡C)
your samples with liquid nitrogen after collecting
and avoid thawing to prevent RNA degradation.
Cell sample: After collecting, wash more than
one time with cold PBS and make pellet. Then,
freeze your sample rapidly with liquid nitrogen.
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| After GeneFishingTM DEG servc |
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| 1. If I take the DEG analysis service, can I know the gene function as well? |
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We will only provide DEG gene information through NCBI blast search as well as sequence data.
For the further research on the gene function, you should design a personal. |
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2.
After cloning and sequencing, two genes were
identified to be
involved in one
DEG band. Which gene should I select as DEG? |
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It can happen if two genes have very similar sizes. In this case, you should confirm each gene by
RT PCR or real-time PCR method using target primers of each gene. |
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3.
As a result of sequencing, the DEG was identified
as an unknown gene.
What procedure
should I take for the next experiment? |
In general, after confirming DEG with target RT-PCR, a full length cDNA should be made .
Then, you can do various functional study using this cDNA. |
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| 4. If identified DEG band size is shorter than 200bp, is it possible to identify the gene? |
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Although sometimes we mark DEG under 200bp, gene analysis is easier when DEG size is more
than 250bp. |
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