Download M1205-2, -4 96 Well Gel Filtration Plate

FOR RESEARCH USE ONLY!
02/17
96 Well Gel Filtration Plate
(Cat# M1205-2, -4; 2 or 4 Plates; Store at 4ºC)
I.
Introduction:
The 96 Well Gel Filtration Plates are pre-packed with a highly efficient gel filter which is ideal for removing excess unincorporated dye
terminators, freeing nucleotides from sequencing and labeling reactions, protein purification, desalting and buffer exchange. Low molecular
weight material is retained in the gel matrix, facilitating the purification of DNA fragments larger than 20 bases in length. Since 96-Well G50 Gel Filtration Plates are designed to purify DNA fragments >20 bases only, they are not recommended for PCR product primer removal.
II. Application:
 Removing excess unincorporated dye terminators
 Freeing nucleotides from sequencing and labeling reactions
 Protein purification
 Desalting
 Buffer exchange
III. Key Features:
 Purification in only 10 minutes!
 Sample Volume: 20 to 50 μl
 Filtrate: >20 bases
 Filtration Plates are pre-packed with a highly efficient gel filter
IV. Sample Type:
 DNA fragments larger than 20 bases in length
V. Kit Contents (96 Well Gel Filtration Plate):
Components
M1209-2
M1209-4
96 Well Gel Filtration Plate
2 Plates
4 Plates
VI. User Supplied Reagents and Equipment:
 2 ml collection plate
 0.35 ml collection plate
VII. Shipment and Storage:
All the reagents are shipped and stored at room temperature (4ºC) for up to atleast 6 months without showing any reduction in performance.
VIII. Reagent Preparation and Storage Conditions:
 In the event of gel drying/cracking add 50-100 µl of ddH2O to each well of the G-50 Plate prior to use.
IX. Purification/Removal/Desalting Protocol:
Please read the entire instruction manual prior to starting the Protocol Procedure.
1. Plate Preparation

Remove the adhesive film from the G-50 Plate. Place the G-50 Plate on a 2 ml collection plate. Centrifuge at 2,500g for 5 minutes.
2. Sample Loading

Transfer the G-50 Plate to a 0.35 ml collection plate. Carefully load the sample (20-50 µl) onto the center of each gel filter surface.
3. Purification Recovery

Centrifuge at 2,500g for 5 minutes. Each purified sample can be recovered at the bottom of the 0.35 ml collection plate.
(approximately the same volume as the loaded sample).
Buffer Exchange Protocol
1. Plate Preparation

Remove the adhesive film from the G-50 Plate. Place the G-50 Plate on a 2 ml collection plate and centrifuge at 2,500g for 5 minutes.
Discard the flow-through and place the G-50 Plate back on the same 2 ml collection plate.
2. Buffer Loading

Add 350 µl of desired buffer to each well of the G-50 Plate. Centrifuge at 2,500g for 5 minutes.
3. Sample Loading

Transfer the G-50 Plate to a 0.35 ml collection plate. Carefully load the sample (20-50 µl) onto the center of each gel filter surface.
4. Purification Recovery

Centrifuge at 2,500g for 5 minutes. Each purified sample can be recovered at the bottom of the 0.35 ml collection plate (approximately
the same volume as the loaded sample).
X. Functional Test Data:
Test
Original DNA
2
3
DNA Conc
23.70 ng/µl
18.7 ng/µl
16.2 ng/µl
Total DNA
1185 ng
935 ng
810 ng
Recovery
----------78.9 %
87.3%
Figure 1. Gel slice DNA fragments ranging from 250 bp-10 kb were extracted using the Gel Extraction Kit (lane 1,2,3).
The purified DNA from a 50 µl eluate was analyzed by electrophoresis on a 1% agarose gel. M = 1 Kb DNA Ladder (control,
total DNA = 1100 ng).
155 S. Milpitas Blvd., Milpitas, CA 95035 USA | T: (408)493-1800 F: (408)493-1801 | www.biovision.com | [email protected]
FOR RESEARCH USE ONLY!
XI.
02/17
Related Products:
Product Name
Avg DNA Frag Extraction Kit
DNA Cleanup Kit
DNA Cleanup Midi Kit
DNA Cleanup Maxi Kit
Ext Gel/PCR Combo Kit
Gel Extraction Kit
Gel Filtration Columns
Gel Filt. Dye Removal Columns
Mini DNA Frag Extraction Kit
96 Well Gel Extraction Kit
96 Well Gel Filtration Plate
96 Well PCR Cleanup Kit
PCR Cleanup Binding Plate
PCR Cleanup Kit
Trad Gel/PCR Combo Kit
XII.
Cat. No.
K1366-100, -300
K1367-100, -300
K1368-100, -300
K1369-10, -25
K1364-100, -300
K1362-100, -300
M1206-2, -50
M1207-2, -50
K1367-100, -300
K1370-2, -4
M1205-2, -4
K1371-2, -4
M1208-5
K1365-100, -300
K1363-100, -300
Size
100 Preps, 300 Preps
100 Preps, 300 Preps
100 Preps, 300 Preps
10 Preps, 25 Preps
100 Preps, 300 Preps
100 Preps, 300 Preps
2 Pcs, 50 Pcs
2 Pcs, 50 Pcs
100 Preps, 300 Preps
2 Plates, 4 Plates
2 Pcs, 4 Pcs
2 Plates, 4 Plates
5 Plates
100 Preps, 300 Preps
100 Preps, 300 Preps
General Troubleshooting Guide:
Problem
Low RNA Yield
Cause
. Agarose gel did not dissolve completely
. Incomplete Wash Buffer preparation.
. Incorrect DNA elution step
Eluted DNA Does Not Perform Well
in Downstream Applications
. DNA was denatured (a smaller band
appeared on gel analysis).
. Primer dimer contamination in the final
PCR elution product.
Solution
. Ensure the agarose gel was melted/dissolved completely
between 55-60ºC for 10-15 minutes, or until no gel is
visible. If undissolved agarose remains in the sample, the
DFH Column could clog and some DNA will be
unrecoverable. DNA can be denatured if the incubation
temperature exceeds 60ºC. If using more than 300 mg of
agarose gel, separate it into multiple 1.5 ml microcentrifuge
tubes.
. Add absolute ethanol (see the bottle label for volume) to
Wash Buffer then mix by shaking for a few seconds. Check
the box on the bottle. Be sure and close the bottle tightly
after each use to avoid ethanol evaporation.
. Ensure that Elution Buffer, TE or water is added into the
CENTER of the DFH Column matrix and is completely
absorbed. If DNA fragments are larger than 5 kb, use preheated Elution Buffer, TE, or water (60~70ºC). If using
water for elution, ensure the water pH is ≥8.0. ddH2O
should be fresh as ambient CO2 can quickly cause
acidification.
. Ensure the agarose gel was melted/dissolved completely
between 55-60ºC for 10-15 min, or until no gel is visible.
DNA can be denatured if the incubation temperature
exceeds 60ºC. Incubate the eluted DNA at 95ºC for 2
minutes then cool down slowly to reanneal the denatured
DNA.
. Gel purification should be performed if primer dimers are
visible in the agarose gel following PCR reactions. Simply
cut the PCR product from the gel and avoid the primer
dimer. Using an additional 80% ethanol wash will reduce
primer dimer contamination when performing PCR
cleanup.
FOR RESEARCH USE ONLY! Not to be used on humans.
155 S. Milpitas Blvd., Milpitas, CA 95035 USA | T: (408)493-1800 F: (408)493-1801 | www.biovision.com | [email protected]