LightCycler Capillaries

LightCycler Capillaries from optical Polycarbonate NOW REDUCED IN PRICE. A clever alternative to the Glass-capillary

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Total RNA purification kit	159.00 Euro
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Your advantages:

- no risk of breaking

- no risk of contamination

- same sensitivity

- no or less change in protocol

- about 40 to 50 % better price

- easy post sample recovery

Catalog-no:    Description Amount    Euro             Shop

A31-960     LightCycler qPCR-capillaries: in racks with Caps 10x96
~~270.00~~
245.00
add

A31-30SKO
LightCycler Starter-Set I : Carousel-Adapter for LightCycler 1,2 or 1,5 and 1 rack, NEW transferpin and cap 1 Rotor + 1x96 + Transferpin 320.00 add

A31-30SK1 LightCycler Starter-Set II: Carousel-Adapter for LightCycler 2 and 1 rack, NEW transferpin and cap 1 Rotor + 1x96 + Transferpin 320.00 add

A31-30TP Transfer Pin 1 pcs 75.00 add

LightCycler Capillaries for RocheLight Cycler Deutsche Beschreibung
Light Cycler Capillaries: Order by E-Mail

NEW Transfer-Pin:
For more comfortable handling of the capillaries and the caps a special transfer pin can be used. With this transfer pin you can take and hold the caps as well as the capillaries handle them as usual, but more comfortable

Short Protocol:

- Take your existing protocol for glass-capillaries from LightCycler

- Prepare your existing PCR-Mastermix. The reaction volume is from 15 – 50 µl
NOTE: The PE capillaries do not require any use of BSA in the reaction mix. Precipitates from BSA can decline the optical density of PE-capillaries.

- Wear hand cloves and use the transfer pin for easier handling

- Take the capillaries out the rack and pipette the PCR-Master mix and the other

components into the upper part of the PC-Capillary

- For an easier handling may you put the PC-Capillary in a 1,5 ml reaction tube using a

standard PCR rack

- Either you transfer the capillary with the 1,5 ml tube to a table top centrifuge (2-3

seconds) OR with a short jerk you bring the Mix down to the bottom.

- Note: Do not use the carousel-adapter from GeneON for any centrifuge system. It is

not constructed for high speed centrifugation. GeneOn is not liable for any damage

that occurs for the wrong usage of the carousel in a centrifuge

- Close the capillary with the cap using the transfer pin

- Ensure that the cap close tightly and transfer the capillary the carousel (GeneOn

Rotoradapter for Lightcycler)

PCR Program

Because of the larger diameter and the more thick walls of the PC-capillary it might be necessary to increase the “hold-times” for denaturation and annealing. We recommend increasing the times with about 10 seconds. If you already use longer times the maximum prolongation is 5 seconds. Depending on you GC-content it might be better be increase the denaturing temperature from 95°C to 97°C for higher efficiency.

Examples for PCR-program

Denaturing times at 95°C
	glass	PC-Cap..	glass	PC-Cap.	glass	PC-Cap.
Denaturing	0 sec	10 sec	5 sec	10-12 sec	> 7 sec	> 10 sec
Annealing	0 sec	10 sec	5 sec	10-12 sec	> 7 sec	> 10 sec
Extension	10 sec	10 sec	5 sec	10-12 sec	> 7 sec	> 7 s

Denaturing time at 97°C
	glass	PC-Cap.	glass	PC-Cap.	glass	PC-Cap.
Denaturing	0 sec	5 sec	5 sec	7-10 sec	> 7 sec	> 10 sec
Annealing	0 sec	10 sec	5 sec	10 sec	> 7 sec	> 10 sec
Extension	10 sec	10 sec	5 sec	5-8 sec	> 7 sec	> 7 s

Note: Longer denaturing times than 10-11 seconds brings not a better result

FAQ:

Melting curve:
The curve shows higher values or the spread is larger: Decrease the slope of the heating step (< 0.1 increments)

Sample recovery:
- cut the capillary with a pair of scissors
- spin the solution into a reaction tube

and pipette the PCR-product on a gel
or

- remove the cap
- contracting by spinning the PC-Capillary

in to a reaction tube for further steps

Fluorescence detection
You can use all standard dyes for PC-Capillaries of GeneON.
In a few cases ROX dye is brings not the expected results. If you are sure that your PCR was successful (by checking on an agarose-gel) it is impossible to increase the PCR-result with any optimization, unfortunately.

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