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Direct WGA Kit

Cat. No. Amount Price (EUR) Buy / Note
PCR-382S 20 reactions x 20 μl 110,00 Add to Basket/Quote Add to Notepad
PCR-382L 100 reactions x 20 μl 440,00 Add to Basket/Quote Add to Notepad

For in vitro use only!

Shipping: shipped on blue ice

Storage Conditions: store at -20 °C

Shelf Life: 12 months

Applications:

  • Genotype analysis
  • PCR and real-time PCR
  • Construction of genomic library

Description:
Direct WGA Kit is a complete system for whole genome amplification from various tissues or samples directly without DNA purification processes. Very little amount of samples, several miligram or microliter volume, are required for the direct WGA. About 10 μg DNA products could be obtained in a standard reaction. The enzyme mix and buffer system are designed to tolerate against most amplification inhibitors found in crude samples. Phi29 DNA polymerase, the major polymerization enzyme of this kit, isothermally amplifies the genomic DNAs included in the samples with multiple displacement mechanism. Phi29 DNA polymerase could produce DNA strand up to 70 kb long with high fidelity. All required components including enzymes, buffers, dNTPs, random primers, and sample pretreatment reagents are supplied in this kit. The amplified DNA products could be applied for successive PCR, genotyping, and library construction.

  • Fast and uniform amplification across entire genome
  • Multiple Displacement Amplification by Phi29 DNA polymerase
  • Direct WGA from Whole blood, animal tissues, plant leaves and seeds, clinical & forensic sample [Saliva, Buccal swab, Hair root, Blood stain (toilet paper or paper)]

Content:

ComponentPCR-382SPCR-382L
1 M DTT100 μl500 μl
PBS Buffer20 μl100 μl
DB1.0 ml5 x 1.0 ml
NB40 μl200 μl
Primer Mix20 μl100 μl
Enzyme Mix20 μl100 μl
Reaction Buffer240 μl1.2 ml
dNTP Mix (each 10 mM)40 μl200 μl


Preparation Procedure

1. Preparation of DM Buffer

  • for one reaction mix 50 μl DB with 5 μl 1 M DTT (for blood samples mix 5 μl DB with 0.5 μl 1 M DTT)
  • Please note: DM Buffer should be freshly prepared for use


2. Sample Preparation

for Blood Samples

  • Add 1 μl of PBS Buffer to 0.5-1 μl of whole blood sample.
  • Add 1.5 μl of DM Buffer and mix by pipetting.
  • Incubate on ice for 10 min.
  • Add 1.5 μl of NB. Briefly vortex and spin down.


for Animal tissue

  • Transfer 50 μl of DM Buffer into a 1.5 ml microtube.
  • Add a tissue slice size of about 5 mm into the DM buffer. Briefly mix by vortexing and spin down.
  • Incubate at room temperature for 10 min.
  • Transfer 2 μl of the supernatant into a new 1.5 ml microtube.
  • Add 2 μl of NB. Mix by pipetting and spin down.


for Plant Leaves or Seeds

  • Transfer 50 μl of DM Buffer into a 1.5 ml microtube.
  • Add a plant leaf cut size of about 5 mm or several small (<1 mm size) pieces of cracked plant seeds into the DM buffer. Briefly mix by vortexing and spin down.
  • Incubate at room temperature for 10 min.
  • Transfer 2 μl of the supernatant into a new 1.5 ml microtube.
  • Add 2 μl of NB. Mix by pipetting and spin down.


3. Preparation of the mix

component20 μl assay
Reaction Buffer12 μl
dNTP Mix
(10 μM)
2 μl
Primer Mix1 μl
Enzyme Mix1 μl
PCR-grade waterfill up to 20 μl

4. Incubation

  • Incubate at 30 °C for 1.5 hours and inactivate the enzyme at 65 °C for 3 min.
  • Please note: Perform the reaction at a thermal cycler or incubator. Water-bath is not recommendable. For PCR, use 1-2 μl of 10-fold diluted product with distilled water. If the PCR is not successful, it is recommended to use 1-2 μl of undiluted product as PCR template.


5. Storage

  • Store amplified DNA at -20 °C.