For every PCR we need DNA, cDNA, or RNA. Most of the PCRs are carried out on blood as a source of DNA. DNA extraction is expensive and time-consuming as well as being a burden to most labs. GT BLB is a special buffer that has been formulated to ease PCR preparation by avoiding DNA extraction. This special buffer can replace DNA extraction for most PCR applications. Only small amount of blood (1 ul per PCR) with GT BLB (4 ul per blood sample per PCR) is necessary to avoid DNA extraction and save time and money in the lab. We have optimized most GT Multiplex PCR kits to work well with BLB. For other uses, we recommend that the user test and optimize the amount of BLB for each reaction.
GT BLB (Blood Lysis Buffer) is a special buffer to enable the technicians to use blood directly in a PCR avoiding DNA extraction. The diluted blood can be used directly to perform molecular tests on multiplex STR-based kits like GT diagnostics and GT Detector HID kits, AneuSure® range of kits, AZFScreen range of kits, and other multiplex PCR-based kits from Genetek Biopharma
Advantages:
- No special treatment is required
- No DNA extraction is needed
- It's inexpensive and easy to use
- A small amount of blood is needed.
- One drop of blood is enough for 100s of PCR since there is about 50 µl in one drop of blood and each µl of blood gives 5 µl of ready-to-use diluted blood.
How to use:
We recommend the use of 1 µl of peripheral blood with 4 µl of BLB and using 1 µl for each PCR. Please use fresh and keep the rest at 4°C until use, though we recommend using it fresh. This buffer can also be used on frozen blood as well as blood cells lysed due to freeze-thawing.
This buffer has been optimized to be used on AneuSure® and AneuSure® Plus, AneuSure® Extra Plus kits, GT AZFScreen, GT AZFScreen Plus, GT Detector range of HID kits, etc. The PCR program must have a hot start program with at least 5 min of initial denaturation at 95˚C. For others’ kits, the user must test and optimize the PCR condition.
Storage:
The GT BLB should be kept at 4ËšC.Genetek also has amniotic fluid lysis buffer (GT AFLB) and chorionic villus lysis buffer (GT CVLB).
Troubleshooting:
We recommend the use of good-quality DNA as a positive control for the first time. The GT BLB should produce the same type of result if the right amount of blood is used. Sometimes, blood has inhibitors that hamper PCR as seen with DNA extracted from these blood samples. Testing more than 200 samples, we did not notice any inhibition. Do not use more than 2 µl of diluted blood since excess blood may inhibit PCR due to the presence of excess Hem.
