|Applications||100 ml Cultured Cell|
1. Lyse Cultured Cells in Straight PCR Lysis Reagent.
2. Incubate for 45 min at 85°C.
3. PCR genotyping with 1 μl lysates.
★ Time saving : Less hands-on time. Crude tail lysates for PCR.
★ Money saving : Cost-effective reactions.
★ Safe : No organic reagents.
★ Environmental : Less waste (organic reagents, tubes, tips, etc...)
★ Reliable and efficient : Virtually 100% success rate with high yields.
1. Complete lysis：Big tissue clumps should not be observed after digestion. It is recommended to vigorously shake the bottle (containing microfuge tubes) for 2-3 sec anytime, once or twice, after tissues begin to partially dissolve. This will physically disperse partially digested tissues and reposition microfuge tube, in which tails are separated from lysis reagents, thereby facilitating overall lysis efficiency.
2. Proteinase K inactivation：Inactivation of proteinase K by incubating samples at 85°C-86°C for 45-50 min is critical to protect Taq polymerase from proteinase K.
3. Tissue size：The size of tails should be 0.5 cm or slightly smaller. Use a minimal volume (0.5-1 μl for 50 μl PCR reaction) of crude lysates for PCR amplification. Too much Straight PCR Lysis Reagent inhibit PCR efficiency.
4. Small tubes and evaporation：To minimize evaporation, use a 0.75 ml tube when the reagent volume is less than 100 μl.
5. Small samples and dilution：If the required Straight PCR Lysis Reagent volume is less than 50 μl, dilute the reagent by up to 2-fold with water, while maintaining the same concentration of proteinase K. If the Straight PCR Lysis Reagent is '2-fold' diluted, apply '2-fold' more crude lysates for PCR reaction.
6. PCR machine：PCR machines are occasionally a source of technical problems.
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