Direct WGA Kit

Catálogo Nº Apresentação Preço (R$) Comprar
PCR-382S 20 reactions x 20 μlSob demanda Adicionar ao Carrinho
PCR-382L 100 reactions x 20 μlSob demanda Adicionar ao Carrinho

For general laboratory use.

Envio: shipped on gel packs

Condições de armazenamento: store at -20 °C

Validade: 12 months

Formulários:

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

Descrição:
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 genomeMultiple Displacement Amplification by Phi29 DNA polymeraseDirect WGA from Whole blood, animal tissues, plant leaves and seeds, clinical & forensic sample [Saliva, Buccal swab, Hair root, Blood stain (toilet paper or paper)]

Contente:
Component||PCR-382S||PCR-382L
1 M DTT||100 μl||500 μl
PBS Buffer||20 μl||100 μl
DB||1.0 ml||5 x 1.0 ml
NB||40 μl||200 μl
Primer Mix||20 μl||100 μl
Enzyme Mix||20 μl||100 μl
Reaction Buffer||240 μl||1.2 ml
dNTP Mix (each 10 mM)||40 μl||200 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 (