Diffinity Genomics RapidTip Instruction Manual

Difnity Genomics
RapidTipTM User Handbook
Version D (December 2010)

Table of Contents
Kit Contents 3
storage 3
ProduCt use Limitations 4
ProduCt Warranty and satisfaCtion guarantee 4
QuaLity ControL 5
teChniCaL assistanCe 5
safety information (disPosaL) 5
ProduCt sPeCifiCations 6
introduCtion 7
diffinity teChnoLogy PrinCiPLe 8
eQuiPment to be suPPLied by user 9
PiPetting With the raPidtiP 10
raPidtiP PCr PurifiCation ProtoCoLs 11
troubLeshooting guide 13
ordering information 14
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Kit Contents
Cat Number Product
Description
Contents
RT025-008 Difnity RapidTip for
PCR Purication
1 Pack of 8
RapidTips
RT025-048 Difnity RapidTip for
PCR Purication
1 Box of 48
RapidTips
RT025-096 Difnity RapidTip for
PCR Purication
1 Box of 96
RapidTips
RT025-096 * Difnity RapidTip for
PCR Purication
* Multiple box
purchases (3,
12, 24, and 48)
are also available
All Kits come with a User Manual
Storage
Difnity RapidTips should be stored in low humidity
at room temperature (15-25 degrees Celsius). Under
these conditions, Difnity RapidTips can be stored
and used for up to 12 months without reduction in
performance or quality.
Difnity RapidTip Page 3

Product Use Limitations
Difnity RapidTips are intended for research use
only. Difnity RapidTip is designed for purication
of PCR solutions prior to dideoxy (BigDye) reaction
for Sanger Sequencing. RapidTip is compatible with
most universal pipettors. RapidTip has not been
tested for compatibility with detergents, mineral
oil, nor ready-to-load style PCR master mixes that
contain a density increasing compound.
Product Warranty and Satisfaction
Guarantee
Difnity Genomics will guarantee the performance
of Difnity RapidTips for the uses and performance
as described in our product literature. If the product
fails to perform to your satisfaction for any reason
other than misuse, we will replace the product free
of charge or refund the purchase price - whichever
you choose. Difnity Genomics maintains the right
to update or modify the product to enhance its
performance. A copy of our terms and conditions
can be found on our web site.
If you have questions about the product or its
performance, please contact Difnity Genomics
customer service at 1-877-362-1812.
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Quality Control
Each lot of Difnity RapidTips is tested against
predetermined specications to ensure consistent
quality.
Technical Assistance
Call 1-877-362-1812 between 8am and 4pm Eastern
time Monday through Friday or visit our webpage
for help with common questions.
Safety Information (Disposal)
Difnity RapidTips may be disposed in regular lab
trash. For more information, please consult the
material safety data sheet (MSDS).
Difnity RapidTip Page 5

Product Specications
Capacity: PCR Sample
Size
25 +/- 5 microliters
dsDNA Recovery up to 90%
(100bp to 10Kb)
Impurity Removal
(dNTP, primers, primer-
dimers)
up to 90%
(less than 45-50 bp)
Please note that at this time the Difnity RapidTip
does not remove DNA polymerase. Standard DNA
polymerase such as Taq does not typically interfere with
the sequencing amplication (BigDye) reaction.
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Introduction
Difnity Genomics has developed an innovative new
technology that removes undesirable impurities
from PCR reactions leaving you with nothing
but puried DNA! This revolutionary technology
effectively removes dNTPs, primers, and primer-
dimers while providing up to 90% recovery of pure
DNA fragments ranging in length from 100bp to
10Kb. It dramatically reduces the amount of time
to purify and prepare PCR reactions for use in
downstream applications, such as DNA sequencing.
The RapidTip is one-tip, one-step, and one-minute
for PCR purication.
Difnity RapidTip Page 7

Difnity Technology Principle
The Difnity RapidTip contains everything you
need for PCR purication. The tip is lled with our
proprietary adsorption technology, which has a
differential afnity for PCR reaction components.
The impurities are removed from the solution as
it enters the pipette tip. Dispensing the solution
yields puried, high quality DNA ready to use for
downstream applications. You won’t need additional
equipment, reagents, buffers, or enzymes - only a
pipettor!
.5Kb 1Kb 2Kb
L- + - + - + L
Difnity RapidTip Purication has excellent yield. Equal volumes
of untreated (-) and Difnity RapidTip treated (+) PCR product
(.5Kb, 1Kb, 2Kb) were run on a 1% agarose gel alongside
Invitrogen 1Kb Plus Ladder (L) and labeled with SybrSafe.
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unpuried
Difnity RapidTip
Difnity RapidTip purication improves sequence quality.
Sequence chromatograms show improved peak quality and lower
background in Difnity RapidTip puried samples as compared to
unpuried samples.
Equipment to be Supplied by User
• PCR Sample
• Tubes to store puried DNA
• Pipettor - manual or programmable
• Standard pipette tips for liquid transfer
(if necessary)
Difnity RapidTip Page 9

Pipetting with the RapidTip
Careful pipetting is important to achieve effective
purication with the RapidTip. The following
pipetting guidelines will help ensure optimal results:
1. Aspirate slowly to ensure particles are dispersed
for effective mixing.
If using a manual pipettor, aspirate slowly for the
rst several cycles until the particles are completely
dispersed into the PCR sample.
If using an automated pipettor, set to aspirate on
low speed (2 of 10 or equivalent), dispense on
middle speed (5 of 10 or equivalent) and place into
manual mix mode.
2. Maintain continuous contact between the
end of the pipette tip and your sample. Avoid
disrupting the uid column with air bubbles.
3. Maintain clearance between the end of the
pipette tip and the plastic tube or well holding
sample to avoid plugging the tip during sample
mixing.
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RapidTip PCR Purication Protocols
Prepare Samples:
Difnity RapidTip is optimized for 25 uL reaction
volumes but can effectively purify samples from 20-
30 uL.
• For PCR volumes > 30 uL, please aliquot 25
uL into a new tube for purication.
• For PCR volumes < 20 uL, please dilute to 25
uL. This works best with highly concentrated
DNA samples as your sample concentration
will be reduced.
Prepare tips:
Difnity RapidTip contains proprietary particles that
purify a PCR reaction; these particles can adhere to
the pipette tip walls during shipping. For optimal
results, sharply tap the box 2-3 times on a at
surface so that particles are at the bottom of the tips
(near the retainer).
Please note that it is normal to see ne dust-like particles on the
side of the tip. After you tap the box, you should expect to see
about 2mm of white particles above the retainer at the small end
of the pipette tip.
Purify Samples:
1. Program pipettor to aspirate 50 uL.
Difnity RapidTip Page 11

2. Place Difnity RapidTip on pipettor.
Please note that you can use a multi-channel pipettor to mix
more than 1 sample at a time for even higher productivity.
3. Place pipette tip into 25 uL PCR solution. Mix for
60 seconds (approximately 15 aspirate/dispense
cycles).
Please note that pipetting will be slower than normal - wait
for liquid to completely ll the tip to begin the next mix. It is
not necessary to drive liquid completely out of the tip on every
dispense. The particles should mix completely with the solution
and make it appear cloudy.
4. Dispense all solution into a clean tube when
mixing is complete. Use your pipettor’s blowout
mode for maximum liquid recovery.
Your puried PCR Amplicon is now ready for
downstream Sanger amplication.
If you wish to conrm purication, run some
unpuried and puried sample on a gel in adjacent
lanes to conrm the amplicon band.
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Troubleshooting Guide
Problem Things to try:
Low DNA recovery Run some of the untreated and RapidTip
puried PCR on a gel to check your
amplicon band intensity.
Note: Purify at least 20 uL of your PCR reaction
to optimize DNA recovery.
Slow or difcult
aspiration
Check that pipettor volume is 50 uL
Check that tips are rmly attached to the
pipettor
Note: Difnity RapidTip is incompatible
with detergents, mineral oil, and ready load
PCR mixes that contain density increasing
compounds.
Fluid remains in the
RapidTip
It is normal for a small amount of liquid
(~ 5 uL) to remain inside the Difnity
RapidTip.
Note: Overdispense or blowout all uid on the
last dispense cycle.
Failure to remove
impurities
Verify that particles are at the bottom of
the tip prior to sample treatment; the
pipettor is operating at the proper (slow)
speed settings; and that particles are
effectively mixing with the sample during
pipetting.
Note: Remove trapped air bubbles by tapping
the tip or mix for additional aspirate/dispense
cycles.
Difnity RapidTip Page 13

Ordering Information
You can place orders by phone or on our website.
Difnity Customer Service: 1-877-362-1812
Website: www.difnitygenomics.com/order.asp
For more information visit our webpage:
www.difnitygenomics.com or call 1-877-362-1812
(c) 2009 Difnity Genomics, Inc. All rights reserved. Difnity
Genomics, Difnity, and RapidTip are trademarks of Difnity
Genomics, Inc. All other names may be trademarks or registered
trademarks of their respective owners.
Printed in the United States
Difnity Genomics, Inc. 150 Lucius Gordon Drive, West Henrietta, NY 14586
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Notes:
Difnity RapidTip Page 15

Difnity Genomics RapidTip User Handbook
Version D (December 2010)