Name: 植物细胞基因组DNA提取和扩增试剂盒
Brand: ACTGene Model: 植物细胞基因组DNA提取和扩增试剂盒
Orginal: TW
Price: 0 Quantity: 2
Promotion: No
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IntroductionPlant Cell DNA Extraction & Amplification Kit
Storage : The kit should be stored at -20℃.
Procedure :
All steps are performed at room temperature unless otherwise noted.
A. Plant Cell DNA Extraction
1. Rinse the paper punch and forceps in 70% ethanol prior to use and between the handling of different samples.
2. Punch a 0.5 to 0.7 cm disk of leaf tissue or frozen leaf tissue into a 1.5 ml or 2 ml microcentrifuge tube using a standard one-hole paper punch.
3. Pipette 100ul of Extraction Solution into the tube and vortex briefly.
4. Incubate samples at 95 ℃ for 10 minutes.
5. Add 80ul of Neutralization Solution and 20ul of Preparation Solution to sample and mix by vortexing.
Note: If several extractions will be performed, sufficient volumes of Neutralization Solution and Preparation Solution may be mixed in a ratio of 4:1 before use.
6. Use immediately in PCR or store the neutralized leaf extract at 4℃. Continue with Section B, Step 1.
Note: Prior to storage, remove the undigested leaf tissue by centrifugation and transfer the extracts to new tubes or wells.
B. PCR Amplification
Taq mastermix, Hot-Start Taq mastermix, Taq Blue mastermix, or Hot-Start Taq Blue mastermix can be used for the following PCR amplification steps. Typical final primer concentrations are approximately 0.4uM each. Primer concentrations and thermal cycling parameters should be optimized depending on the system.
1. Add 10ul of PCR mastermix (2x), 4ul of leaf disk extract, and proper amount of primers to a thin-walled PCR microcentrifuge tube or plate. Bring the total volume to 20ul with PCR grade water. Gently mix well.
Note: If less than 4ul of leaf disk extract is added to the PCR reaction volume, use a 50:50 mixture of Extraction Solution : Neutralization Solution to compensate the volume of tissue extract to 4ul.
2. Perform thermal cycling using optimized amplification parameters.
3. Load the amplified DNA onto an agarose gel after the PCR is completed.
Kit Specification:
Reagents 10-rxn 100-rxn 1000-rxn
Extraction Solution 3ml in 5ml bottle 24ml in 30ml bottle 250ml in 250ml or
2x125ml bottles
Preparation Solution 0.35ml in 1.5 ml vial 3ml in 5ml bottle 30ml in 30ml bottle
Neutralization Solution 3ml in 5ml bottle 24ml in 30ml bottle 250ml in 250ml or
2x125ml bottles
Hot-Start Taq Blue Mastermix 0.15ml in 1.5ml vial 1.2ml in 1.5ml vial 12ml in 15ml conical tube