AIM: The purpose of the exercise is to carry a PCR reaction with the DNA templates in order to determine human sex using PCR. The procedure is based on the amplification of a human amelogenin gene fragment. This gene is shorter by 189bp on chromosome Y as compared with chromosome X, which allows for the determination of human sex by means of PCR. The expected PCR products are 977bp and 788bp long for males, or only 977bp for females.

The kit is designed to perform experiments during two laboratory classes (I – PCR set-up – 1.5 hours; II – electrophoresis – 1.5-2 hours). We suggest to perform laboratory experiments in six groups of two students.

EasyPCR XY kit components (1 exercise, DY10A):

6 sets of PCR reagents:

• 6 tubes with 10x reaction buffer (10 reactions per tube)
• 6 tubes with MgCl₂ solution (10 reactions per tube)
• 6 tubes with ready-to-use dNTPs mixture (10 reactions per tube)
• 6 tubes with FORWARD primer (10 reactions per tube)
• 6 tubes with REVERSE primer (10 reactions per tube)
• 6 tubes with PCR – grade water
• 6 tubes with thermostable DNA polymerase (10 reactions per tube)
• 6 tubes with Positive controls (5 reactions per tube) with MALE and FEMALE genomic DNA
• 12 tubes with human genomic DNA for analysis (each for 3 reactions)

Moreover:

• 7 laboratory protocols (one with additional information for a teacher)
• Agarose (5 g)
• Buffer for agarose electrophoresis – 50x TAE (50 ml)
• Ready-to-use M100-1000 DNA Ladder for 10 electrophoretic lanes
• DNA Gel Loading Buffer – 6x GREEN (0.5 ml)

EasyPCR XY kit components (5 exercises, DY105A):

6 sets of PCR reagents:

• 6 tubes with 10x reaction buffer (50 reactions per tube)
• 6 tubes with MgCl₂ solution (50 reactions per tube)
• 6 tubes with ready-to-use dNTPs mixture (50 reactions per tube)
• 6 tubes with FORWARD primer (50 reactions per tube)
• 6 tubes with REVERSE primer (50 reactions per tube)
• 6 tubes with PCR – grade water
• 6 tubes with thermostable DNA polymerase (50 reactions per tube)
• 6 tubes with Positive controls (25 reactions per tube) with MALE and FEMALE genomic DNA
• 12 tubes with human genomic DNA for analysis (each for 15 reactions)

Moreover:

• 7 laboratory protocols (one with additional information for a teacher)
• Agarose (25 g)
• Buffer for agarose electrophoresis – 50x TAE (100 ml)
• Ready-to-use M100-1000 DNA Ladder for 50 electrophoretic lanes
• DNA Gel Loading Buffer – 6x GREEN (1 ml)