Direct WGA Kit

For general laboratory use.

Shipping: shipped on gel packs

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:

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. Add followings and mix well to make final 20 μl reaction mixture

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.

Component DB contains dangerous substance

Signal word: Danger

Hazard statements
H302 Harmful if swallowed.
H314 Causes severe skin burns and eye damage.
H412 Harmful to aquatic life with long lasting effects.

Precautionary statements
P260 Do not breathe dust/fume/gas/mist/vapours/spray.
P301 + P312 IF SWALLOWED: Call a POISON CENTER/doctor/.../ if you feel unwell.
P301 + P330 + P331 IF SWALLOWED: rinse mouth. Do NOT induce vomiting.
P303 + P361 + P353 IF ON SKIN (or hair): Take off immediately all contaminated clothing. Rinse skin with water [or shower].
P304 + P340 IF INHALED: Remove person to fresh air and keep comfortable for breathing.
P305 + P351 + P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.

For further information see Material Safety Data Sheet.

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