| Questions |
Answers |
| a. |
How should I store the kit? |
These Detection Kits 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. |
| f. |
How many reactions does each sample need? |
Only one independent PCR reactions should be conducted simultaneously for each DNA sample. Four control reactions (three for positives and one for negative) should be companied on each run. |
| g. |
What dyes do these kits use? |
FAM and VIC fluorescence. |
| h. |
What is the detection method of these Detection Kits? |
This is one kind of SNP genotyping methods, and we use two specific probes labeled with different fluorescence to detect a SNP site. |
| i. |
How do I determine the genotype of sample? |
You can determine the genotype of sample the same as the positive control nearest to it in the Allelic Discrimination map. |
| j. |
Can I use Post Read function only? |
You can perform the test without Pre Read.
However, It is strongly recommended to use Pre Read to avoid the interference from background noise if your instrument has this function. |
| k. |
Should Positive controls have fluorescent signal? |
Positive controls should have strong fluorescent signals and can show four obvious regions with NTC on Allelic Discrimination map. |
| l. |
Should each sample have fluorescent signal of SNP? |
Each sample has its own SNP types which should have fluorescent signal. If there are some interferents in samples, the locations of signal may move toward the origin (NTC site). |
| m. |
Do I need to duplicate my tests? |
Based on the description on package insert, duplication is not always needed. It needs to be conducted according to the QC guidance of each lab. |
| n. |
Does the NTC show obvious fluorescent signal? |
NTC is a control without DNA sample, and it may still have very weak signal, which has no effect on Allelic Discrimination. High fluorescent signal shown represents an artificial mistake and has sample DNA contamination(s). |
| o. |
How do I set up the software program of the quantitative PCR instrument? |
PharmiGene provides program setting instructions for a few quantitative PCR detection systems. Please do not hesitate to contact us for the further information if needed. |
| p. |
How do I contact the Technical Support dpartment of PharmiGene? |
Please call +886-2-26959800 or email service@pharmigene.com to request a PGI’s service. |
