GV-High-Efficiency DNA Purification Kit 50T

Cat No:GV-PCR-P-50

This kit can recover DNA fragments and exclude impurities such as salt, protein, RNA. For longer than 500bp fragments, the recovery is above 90%.For 200 bp-500bpfragments, the recovery is above 70%.For 80 bp-200bpfragments, the recovery is about 60%. The Kit can recoversingle, double chain and ring plasmid DNA fragments.

The kit apply an unique balance liquid treated adsorption membrane, a reagent that can activate the silicon substrate membrane will improve adsorption ability and enhance uniform and stability of the column. The membrane can specially absorb the DNA.It can also eliminate the bad environmental factors such as heat, moisture to adsorption column.

 

Kit Components, Storage, Stability

Component

Amount

Attention

Buffer PCR-GA

60 ml

 

Buffer PCR-BL

12ml

 

Wash buffer

13 ml

Add 52ml ethanol before use

Elution

4 ml

 

Spin Columns

50

Max adsorption up to 20μg each

1.5ml centrifuge tube

50

 

Handbook

1

 

 

 

 

 

Buffer PCR-GA : DNA Binding Buffer, airtight storage at room temperature.

Buffer PCR-BL : Equilibrium Buffer. Equilibrium solution can activate silicon substrate membrane and improve adsorption ability, uniform and stability of the column. It makes the membranesingle adsorb the DNA. It can also eliminate the bad environmental factors such as heat, moisture to adsorption column

Wash Buffer : Desalting Buffer. Add ethanol as the volume marked on bottle label before use, mix completely. Airtight stored at room temperature.

Elution:10mM Tris-HCl, pH8.0. Airtight stored at room temperature.

 

Announcements

1. DNA store at 10 mM Tris-HCl, pH8.0.

2. Use the column treated byequilibrium buffer the very day, don’t storage for long time.

3.Buffer PCR-BL contains irritant compounds, wear latex gloves and glasses to avoid contaminating skin, eyes and clothes, beware of suction to nose. If contaminate skin or eyes, please wash with plenty of water or saline immediately,seeing the doctor for advice when necessary.

 

Preparation reagent

1.     Prepare Tip tube, centrifuge tube without nucleic acids or nucleic acid enzyme pollution.

2.     Add 52ml ethanol to Wash Buffer before use.

3.     If theBuffer PCR-GAhas precipitate, redissolve by warming to 37℃ and cool to room temperature before use.

4.   Warm the elution or ddH2O to 65℃ to improve elution efficiency.

 

Procedure

1.  Add three times volume of Buffer PCR-GA into PCR product, complement the volume to 50ul with TE Buffer if the volume of PCR products is less than 50μl, mix the solution well.

Note: For DNA fragment less than 200bp, add Buffer PCR-GA with the ratio of 1:5.

2. Deal the centrifugal column before the supernatant banding column: add 200ul Buffer BL to the column, discard Equilibrium Buffer in collection tube.

3. Add the solution from Step 1 in centrifugal column, and stand for 2 minutes, centrifuge for 30s at 12,000rpm. Add twice if the volume is too large to be added once, discard the flow-through.

4.  Add 500μl Wash buffer (add 52ml ethanol before use) to the Spin Columns, and stand for 2 minutes, centrifuge for 30s at 12,000rpm, discard the flow-through.

5. Repeat step 4.

6. Centrifuge for 1 minute at 12,000rpm to remove residual ethanol.

7. Transfer the column to a new sterile 1.5ml micro centrifuge tube, place the centrifugal tube open for 5 ~ 10 mins to volatilize ethanol.

8. Add 2060μl Elution (pre-warm to 65℃), and stand for 2 minutes.

9. Centrifuge for 2 minutes at 12,000rpm, purified DNA Lie in the solution at tube bottom, store at -20℃.

 

Detection

PCR products purified by the Kit can be detected by agarose gel electrophoresis and ultraviolet specrophotometer to test the concentration and purity.

DNA hasabsorption peak at 260nm. Make OD260 valuebetween 0.1 and 1.0 during testing. OD260=1 is equivalent to about 50 μg/ml double-stranded DNA, 40 μ g/ml single DNA.

Nucleic acid concentration(μg/ml)=OD260×50×dilution ratio.

OD260/OD280 value should be 1.7~2.0.

 

Troubleshooting

Problem

Cause

Solution

Low recovery or no band

Have not add ethanol into Wash buffer

Add 52ml ethanol into Wash Buffer

Misuse the Elution buffer

Use Elution offered by the Kit

Elution don't fully

Make sure enough elution time and pre-warm Elution to 65℃

High pH of electrophoresis buffer

Use fresh buffer electrophoresis to test sample

Little sample, low concentration

Increase the volume of sample

Purified DNA is not suitable for experiment

Ethanol residue

Extendairing time appropriately when the room temperaturelow.Put it in 37 ℃incubator for drying.

Salt residue

Make sure washing dosage and washing times is enough.

 

 

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