PCR labeling DIG Probe kit
Cat number: 001.1145
Size:25T
Storge:-20 ℃
The labeled probe application：
FISH, ISH,
Southern blotting, Northern blotting,
Bacteria ISH.
1. Introduction:
PCR labeling DIG Probe kit is one rapid method for preparing DIG-labeled Probe. When the template is available in not too long (<2kb). It requires no optimization than other methods and produces a high yield of DIG probe. In PCR labeling, DIG-dUTP was add into PCR specific region of the template DNA. The result is a highly labeled, very specific, and sensitive hybridization probe.
2.PCR DIG Probe Synthesis Kit
1) Enzyme mix,
2) PCR DIG probe synthesis mix, 10×.
3) PCR buffer with MgCl2, 10×. [1]
4) dNTP stock solution, 10×. [1]
3.Prepare DNA and primers:
(1)DNA 50ng
(2)Forwards primer:0.5uM solution
(3) Reveres primer:0.5uM solution
4. Prepare PCR reaction system:
PCR buffer with MgCl2, 10×: 5ul
PCR DIG mix,10×. 5ul
Forwards primer: 1ul
Reveres primer: 1ul
Enzyme mix: 1ul
Template DNA: (10-20ng) 1-2ul
Sterile H2O: Add up to 50ul
Mix 1 min
5.Set up PCR system:
1) Initial denaturation: 95°C 3 min –
2) 95°C 30 s, 60°C 30 s, 72°C 40 s
Cycles:1 – 10
95°C 30 s, 60°C 30 s, 72°C 40 s
Cycles: 11 – 30
Final elongation 72°C 7 min
6. Next application:
1) Use 5-8 μl of the reaction mixture to check the results of the labeling reaction.
2) PCR productions are stored at 4°C.
3) PCR-generated probes can be used directly in the hybridization reaction. It not require clean up.
4) 2 μl/ml in hybridization solution is suitable.
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