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PreCR® Repair Mix
Catalog #
M0309S
M0309L
Size
30 reactions
150 reactions
Concentration
Categories: DNA Manipulation, DNA Manipulation, DNA Repair Proteins | More +
Applications: DNA Repair, PCR, PCR
Product
Information
FAQs &
Tech Tips
Description
Properties and Usage
Notes
Protocols &
Manuals
Other Tools &
Resources
Quality &
Safety
Legal
Information
Advantages and Features
Related Products
References
Description
The PreCR® Repair Mix is an enzyme cocktail formulated to repair damaged template DNA prior to its use in the polymerase chain reaction
(PCR), microarrays or other DNA technologies. PreCR is active on a broad range of DNA damages, including those that block PCR (e.g.
apurinic/apyrimidinic sites, thymine dimers, nicks and gaps) and those that are mutagenic (e.g. deaminated cytosine and 8-oxo-guanine). In
addition, it will remove a variety of moieties from the 3´end of DNA leaving a hydroxyl group. The PreCR Repair Mix will not repair all
damages that inhibit/interfere with PCR. For example, it will not repair 8-oxo-7,8-dihydro-2'deoxyadenosines or fragmented DNA. In fact, the
ligase present in the mix is very active at sealing nicks in DNA but does not ligate blunt ends or nicks near a mismatch effectively. This
limits the possibility of chimeric gene formation as a product of ligation. The PreCR Repair Mix can be used in conjunction with any
thermophilic polymerase.
DNA is susceptible to many types of damage resulting from exposure to a variety of chemical or environmental agents, manipulation or
simply aging. The DNA Damage and PreCR Table lists some possible DNA samples and the impact of the damage they may undergo.
Note: The extent of damage caused by exposure to different reagents can vary, and its importance will depend on how the DNA is being
used.
Table 1:
Types of DNA Damage
Figure 1: Repair of different types of DNA damage with the PreCR Repair Mix.
The gel shows trial amplifications from damaged DNA that was either not treated (-) or treated (+) with the PreCR Repair Mix. Type of DNA
damage is shown. Note: heat treated DNA is incubated at 99°C for 3 minutes. Marker M is the 2-Log DNA Ladder (NEB #N3200 ).
Highlights
Suitable for PCR reactions, microarrays or other DNA technologies
Easy-to-use protocols
Does not harm template
Kit Components
The following reagents are supplied with this product:
ThermoPol® Reaction Buffer Pack
Store at (°C)
Concentration
-20
10X
L1 Primer Mix
20 μM
UV damaged Lambda DNA
0.5 ng/μl
β-Nicotinamide adenine dinucleotide (NAD+)
-20
PreCR® Repair Mix
Purified BSA
Advantages and Features
100X
1X
-20
10X
Applications
Repair of DNA prior to its use as a template in PCR or other DNA technologies.
Properties and Usage
Related Products
Companion Products
Taq 2X Master Mix
Taq DNA Ligase
Taq DNA Polymerase with ThermoPol® Buffer
TriDye™ 2-Log DNA Ladder (0.1 - 10.0 kb)
Endonuclease IV
Endonuclease VIII
Fpg
Quick-Load® 2-Log DNA Ladder(0.1-10.0 kb)
T4 PDG (T4 Endonuclease V)
Uracil-DNA Glycosylase (UDG)
Bst DNA Polymerase, Full Length
Materials Sold Separately
β-Nicotinamide adenine dinucleotide (NAD+)
ThermoPol® Reaction Buffer Pack
Notes
1. * PreCR Repair Mix components: Taq DNA Ligase, Endonuclease IV, Bst DNA Polymerase, Fpg, Uracil-DNA Glycosylase (UDG),
T4 PDG (T4 Endonuclease V) and Endonuclease VIII.
2. Reaction Conditions: DNA template, 100 µM dNTPs, 1X ThermoPol Reaction Buffer, 1X NAD+ and 1 µl PreCR Repair Mix in a total
reaction volume of 50 µl. Incubate at 37°C.
3. 1X NAD+ Solution: 0.5 mM NAD+.
4. dNTPs are not included with the PreCR Repair Mix.
References
1. Diegoli TM, Farr M, Cromartie C, Coble MD, Bille TW. (2012). An optimized protocol for forensic application of the PreCR™ Repair Mix
to multiplex STR amplification of UV-damaged DNA.. Forensic Sci Int Genet. Jul, 498-503. PubMedID: 22001155
FAQs
FAQs
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Will PreCR ligate my DNA fragments?
How much DNA will PreCR repair?
What are the liquid volumes of the PreCR Repair Mix Components?
Will treating my DNA with the PreCR Repair Mix hurt my reaction?
What is the sequence of the L1 primer mix?
How is the damaged DNA that you test the PreCR Repair Mix against created? Can I buy this damaged DNA separately?
Why does my control reaction not give a detectable amplicon?
Can I buy any of the PreCR Repair Mix components separately?
Will the PreCR Repair Mix blunt the ends of the DNA?
Why don’t I see the expected band from my repaired template?
Is my buffer compatible and which reaction should I use?
Does the PreCr Repair Mix insert random nucleotides into the sequence that it repairs?
Does the PreCR Repair Mix contain any contaminating human DNA? What are the quality controls that are used to test that?
Does the PreCR Repair Mix remove covalent modifications from DNA bases, such as biotin or digoxigenin? Does it repair
mismatches/ extra bases in DNA?
If there are gaps and nicks remaining from a ligation reaction, will the PreCR Repair Mix repair all of these, so that the DNA
will be suitable for microinjection into mice, for example?
Can the PreCR Repair Mix be used for paraffin-embedded DNA?
The repaired DNA will be used for an Nsp1 or Sty1 digestion followed by an adapter ligation, and PCR. Do you recommend
cleanup of the PreCR Repair Mix reaction prior to this process?
Can the PreCR Repair Mix repair damage in both single and double stranded DNA? Or, does it require a double stranded DNA
as a template?
Is the addition of dNTPs necessary for the PreCR Repair Mix to work properly?
If I had a DNA template with mutation sites (ie. 8-oxoguanine or deaminated cytosines) that are directly adjacent to each other
on opposite strands would treatment with PreCR™ Repair Mix cause a double strand nick/break?
21. What gap lengths can be repaired with the PreCR Repair Mix?
22. Does the PreCR Repair Mix work with less concentrated amounts of DNA (e.g. 500pg-1ng) than the amounts recommended?
23. When doing bisulfite treatments of templates, the subsequent PCR reactions can pose a problem as the DNA seems to be
very labile after the treatment. Can the PreCR Repair Mix improve these PCR results?
24. Is it necessary to clean the PreCR reaction prior to carrying out quantitative PCR?
25. What other components may be necessary that are not supplied with the PreCR Reaction Mix?
26. When working with fragments, will the ends be ligated together by the PreCR Reaction Mix?
27. Can T4 DNA ligase be used to ligate across an abasic site?
28. How are abasic sites repaired by the PreCR Reaction Mix? Are new nucleotides put in and ligated? Is the existing nucleotide
repaired?
Protocols
Datacards
Protocols
1. Sequential Reaction Protocol for PreCR Repair Mix
2. Standard Reaction Protocol for PreCR Repair Mix
3. Control Reaction Protocol for PreCR Repair Mix
Datacards
The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and
quality controls. The following file naming structure is used to name the majority of these document files: [Catalog
Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the
Technical Support Form for appropriate document.
M0309Datasheet-Lot0051205
M0309Datasheet-Lot0091210
M0309Datasheet-Lot0091303
M0309Datasheet-Lot0091308
M0309Datasheet-Lot0101308
M0309Datasheet-Lot0101402
M0309Datasheet-Lot0101407
M0309Datasheet-Lot0101410
M0309Datasheet-Lot0101501
M0309Datasheet-Lot0111507
M0309Datasheet-Lot0121512
M0309Datasheet-Lot0121609
M0309Datasheet-Lot0121703
M0309Datasheet-Lot0131703
Usage Guidelines & Tips
Troubleshooting Guides
Usage Guidelines & Tips
DNA Damage and PreCR
Troubleshooting Guides
PCR Troubleshooting Guide
Quality Control
Datacards
Safety Data Sheet
Quality Control
Quality Assurance Statement
The PreCR repair mix was tested for the ability to rescue successful amplification from UV damaged Lambda DNA (NEB
#N3011). The mix was also tested for the ability to cleave oligonucleotide substrates containing thymine glycol, deoxyuracil, or
8-oxo-guanine in the absence of dNTPs. The absence of dNTPs prevents full repair and allows the presence of specific repairtargeting activities to be evaluated.
Safety Data Sheet
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
PreCR® Repair Mix
ThermoPol® Reaction Buffer Pack
L1 Primer Mix
UV damaged Lambda DNA
β-Nicotinamide adenine dinucleotide (NAD+)
PreCR® Repair Mix
Purified BSA
Datacards
The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and
quality controls. The following file naming structure is used to name the majority of these document files: [Catalog
Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the
Technical Support Form for appropriate document.
M0309Datasheet-Lot0051205
M0309Datasheet-Lot0091210
M0309Datasheet-Lot0091303
M0309Datasheet-Lot0091308
M0309Datasheet-Lot0101308
M0309Datasheet-Lot0101402
M0309Datasheet-Lot0101407
M0309Datasheet-Lot0101410
M0309Datasheet-Lot0101501
M0309Datasheet-Lot0111507
M0309Datasheet-Lot0121512
M0309Datasheet-Lot0121609
M0309Datasheet-Lot0121703
M0309Datasheet-Lot0131703
Legal and Disclaimers
Legal and Disclaimers
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc
(NEB).
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain
additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes
in humans or animals.