EasyPCR HIV + DNA isolation

Index Size
DY25 1 exercise
DY255 5 exercises

Product Features

EasyPCR HIV - DNA isolation and determination of HIV resistance using PCR (+ DNA extraction).

 

AIM: The purpose of the exercise is to carry out a genomic DNA isolation from buccal swabs and a PCR reaction with the isolated DNA templates in order to detect the mutation in the ccr5 gene which confers resistance to HIV in humans. The detection is based on an amplification of ccr5gene fragments:

- 220 bp in homozygotes without the mutation

- 188 bp in homozygotes with the mutation

- 220 and 188 bp in heterozygote with the mutation

 

The kit is designed to perform experiments during two or three laboratory classes (I - DNA extraction and PCR set-up - 2-2.5 hours; II - electrophoresis - 1.5-2 hours or I - DNA extraction - 1-1.5 hours; II - PCR set-up - 1-1.5 hours; III - electrophoresis - 1.5-2 hours). We suggest to perform laboratory experiments in six groups of two students.

 

EasyPCR HIV kit components (1 exercise, DY25)

A) 6 sets of reagents for  DNA isolation from buccal swabs

(each set for 4 isolations)

B) 6 sets of reagents for PCR:  

- 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 control gene: 220 bp fragment (5 reactions per tube)

- 6 tubes with Positive control gene: 188 bp fragment (5 reactions per tube)

C) Moreover:

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

- Agarose (5 g)

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

- Ready-to-use M100-500 DNA Ladder for 10 electrophoretic lanes

- 6x GREEN DNA Gel Loading Buffer (0.5 ml)

 

EasyPCR HIV kit components (5 exercises, DY255):

A) 6 sets of reagents for  DNA isolation from buccal swabs

(each set for 20 isolations)

B) 6 sets of reagents for PCR:  

- 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 control gene: 220 bp fragment (25 reactions per tube)

- 6 tubes with Positive control gene: 188 bp fragment (25 reactions per tube)

C) Moreover:

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

- Agarose (25 g)

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

- Ready-to-use M100-500 DNA Ladder for 50 electrophoretic lanes

- 6x GREEN DNA Gel Loading Buffer (1 m)

 

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

- Automatic pipettes with appropriate tips

- PCR tubes and 1.5 ml Eppendorf tubes

- Sterile swab sticks

- Agarose electrophoresis equipment (apparatus, power supply)

- Microcentrifuge
- Thermoblock, incubator or thermostatic water bath

- UV transilluminator or lamp

- Thermal cycler

- Microwave oven or laboratory burner

- Ethidium bromide solution or other stain for DNA visualization

- Nitrile gloves

 

Shipping conditions

Shipping on dry or blue ice.


Manufacturer: BLIRT S.A.