| Questions |
Answers |
| a. |
How should I store the kit? |
The PG1502 FastGel Kit should always be kept at -20°C in the dark and thawed on the ice while using. |
| b. |
Can I use blood collection tubes containing Heparin as anticoagulant? |
It is highly recommended to use blood collection tubes containing either sodium citrate or EDTA as the anticoagulants. Heparin may have an inhibitory effect on PCR. |
| c. |
Are there preferable DNA extraction kits for extraction gDNA? |
Qiagen QIAamp® DNA Blood Mini Kit is recommended. |
| d. |
What is the required quality of gDNA? |
The OD260/280 ratio of gDNA should be between 1.7 and 2.0. |
| e. |
What is the appropriate amount of DNA in the reaction? |
The appropriate amount of gDNA should be between 25-100 ng/reaction. Inappropriate amount of gDNA, over 100 or less than 25 ng/rxn, may cause weaker band of internal control. |
| f. |
How many reactions does each sample need? |
Only one independent PCR reaction should be conducted for each DNA sample. |
| g. |
Why does the amplification only need two steps? |
We integrate Annealing and Extension steps into one for saving time. |
| h. |
Why the annealing temperature needs to be set up for higher? |
At this temperature, HLA-B*1502 can be distinguished from over 1800 HLA-B* types. |
| i. |
What is the buffer range of temperature during annealing? |
For getting the best performance, the annealing temperature should be kept within +/- 0.5°C. PCR machine usually has steady performance under formal maintenance. |
| j. |
How can I ensure my PCR machine fits the temperature requirement? |
Different PCR machines have various temperature control designs. Upon inquiries, we will provide a testing panel for our customers to adjust their system settings. Please contact our sales department. |
| k. |
How do I distinguish HLA-B*1502 positives from negatives? |
Each human sample has its own internal control signal. Negatives will have only one signal (internal control signal = 250bp). Positives have two signals (419 and 250bp). Please follow the description in package insert. |
| l. |
Does internal control always have signal? |
Internal control is similar to a housekeeping gene. Therefore, it should have signal in each specimen and we can monitor the amount and quality of gDNA sample. |
| m. |
Do I need to duplicate my tests? |
Based on the description in package insert, duplication is not always needed. It needs to be conducted according to the QC guidance of each lab. |
| n. |
Why does the NTC show amplification signal? |
NTC is a control without DNA sample. Ct value shown represents an artificial mistake and has DNA contamination(s) in samples. |
| o. |
How do I contact the Technical Support department of PharmiGene? |
Please call +886-2-26959800 or email service@pharmigene.com to request a PGI’s service. |
