SPEED PCR Conditions Screening Kit

SPEED PCR Conditions Screening Kit

SKU Description Size Price Shipping  
LYM1005 SPEED PCR Conditions Screening Kit 12 cdns x 5 rxns $102 2-5 Days Add To Shopping Cart
LYM1006 SPEED PCR Conditions Screening Kit: Individual Reaction Mix 200 rxns $198 2-5 Days Add To Shopping Cart
LYM1007 SPEED PCR Conditions Screening Kit: with High Fidelity Taq 12 cdns x 5 rxns $102 2-5 Days Add To Shopping Cart
Datasheet:
This Kit contains 12 tubes (Tube 1-12) of 2X PCR mixes. Each mix has a unique buffer composition (Mg2+ concentration, DMSO, PCR Booster, etc). Just simply add your template and primers, and run a PCR. After the tube number giving the best result is identified, you can order that specific tube for further use.

The SKU for specific tube is LYM1006. You will need to provide tube number when you place your order.

This Kit is ideal for identifying the best PCR mix for your routine lab work, such as genotyping of mouse stains.

For high-fidelity amplication applications, please use LYM1007.

Storage

Keep at -20°C for long term storage. Stable at 4°C for three months or for fifteen freeze-thaw cycles.

Protocol

There are 12 different PCR reaction buffers and thus a reaction mix of 14 is generally needed.

                                                              1 reaction                      14 reactions
                Template (DNA)                    1 µl (50-100 ng)             14 µl
                Forward primer (10 µM)        1 µl                                14 µl
                Reverse primer (10 µM)        1 µl                                14 µl
               H2O                                       23 µl                               322 µl


Mix the components well and aliquot 25 µl to each of 12 PCR reaction tubes, then add 25 µl 2X PCR Mix to each of the 12 PCR tubes.

PCR Cycle Profile

As is often the case, this kit is used only when you have difficulties in your PCR. We recommend that you use a touch-down PCR cycle profile to optimize your primer annealing temperature. 

For example, if your primer annealing temperature is around 55°C, use the following PCR cycle program. Adjust accordingly if your primer annealing temperature is higher or lower than 55°C.

Initial DNA denaturation:  94ºC   for 4’ 
  
-  Denature at 94ºC for 30’’ 
-  Anneal at 60ºC for 30’’
-  Extension at 72ºC for 1min/kb
Run 3 cycles

-  Denature at 94ºC for 30’’ 
-  Anneal at 55ºC for 30’’
-  Extension at 72ºC for 1min/kb
Run another 3 cycles

-  Denature at 94ºC for 30’’ 
-  Anneal at 50ºC for 30’’
-  Extension at 72ºC for 1min/kb
Run 25 Cycels

You can also try a gradient touch-down program.

Example Data

(see right)
Other Image