BLIRT EasyPCR I - PCR reaction optimization

Aim: to carry out a series of PCR reactions in order to demonstrate the influence exerted on the efficiency and specificity of the PCR reaction by a change to one of the several parameters being tested.

EasyPCR I - PCR reaction optimization
EasyPCR I - PCR reaction optimization
EasyPCR I - PCR reaction optimization EasyPCR I - PCR reaction optimization
Name Part Number Pack Size
EasyPCR I - PCR reaction optimization DY45 1 exercise
  DY455 5 exercises

 

EasyPCR I – PCR reaction optimization

Aim: presentation of an influence of changing reaction components onto the reaction efficiency. The following will be tested: composition of the reaction buffer, magnesium and nucleotides (dNTPs) concentration  and the amount of template.

The kit is designed to perform experiments during two laboratory classes (I - PCR set-up - 2 hours; II - electrophoresis - 2 hours). We suggest to perform laboratory experiments in six groups of two students.

EasyPCR I kit components (1 exercise, DY45):

6 sets of PCR reagents:

- 5 tubes with 10x reaction buffer (20 reactions per tube)

- 5 tubes with MgCl2 solution (20 reactions per tube)

- 2 tubes with RED polymerases (3 reactions per tube)

- 2 tubes with Master Mix RED (3 reactions per tube)

- 12 tubes with ready-to-use dNTPs mixture (10 reactions per tube)

- 6 tubes with FORWARD primer (20 reactions per tube)

- 6 tubes with REVERSE primer (20 reactions per tube)

- 6 tubes with PCR - grade water

- 6 tubes with thermostable DNA polymerase (20 reactions per tube)

- 6 tubes with DNA template (10 reactions per tube)

Moreover:
- 7 laboratory protocols (one with additional information for a teacher)

- Agarose (5 g)

- 50x TAE buffer for agarose gel electrophoresis (50 ml)

- M100-500 ready-to-use DNA molecular weight standard for 10 electrophoretic lanes
- 6x GREEN DNA Gel Loading Buffer (0.5 ml)

EasyPCR I kit components (5 exercises, DY455):

6 sets of PCR reagents:

- 5 tubes with 10x reaction buffer (100 reactions per tube)

- 5 tubes with MgCl2 solution (100 reactions per tube)

- 2 tubes with RED polymerases (15 reactions per tube)

- 2 tubes with Master Mix RED (15 reactions per tube)

- 12 tubes with ready-to-use dNTPs mixture (50 reactions per tube)

- 6 tubes with FORWARD primer (100 reactions per tube)

- 6 tubes with REVERSE primer (100 reactions per tube)

- 6 tubes with PCR - grade water

- 6 tubes with thermostable DNA polymerase (100 reactions per tube)

- 6 tubes with DNA template (50 reactions per tube)

Moreover:

- 7 laboratory protocols (one with additional information for a teacher)

- Agarose (25 g)

- 50x TAE buffer for agarose gel electrophoresis (100 ml)

- M100-500 ready-to-use DNA molecular weight standard for 50 electrophoretic lanes
- 6x GREEN DNA Gel Loading Buffer (1 ml)

Additional equipment and materials required (not included in the kit):

- Automatic pipettes with appropriate tips

- PCR tubes and 1.5 ml Eppendorf tubes

- UV transilluminator or lamp

- Agarose electrophoresis equipment (apparatus, power supply)

- Microwave oven or laboratory burner

- Thermal cycler

- Ethidium bromide solution or other stain for DNA visualization

- Nitrile gloves

Shipping conditions

Shipping on dry or blue ice.