Molecular
Biology
Handbook &
Selection Guide
c
G-Biosciences 1-800-628-7730 www.GBiosciences.com
Protein Estimation Assays
Lysis Buffers & Systems
Apoptosis Assays
Protein Fractionation Kits
Cytotoxicity Assays
Dialysis (Micro) System
SAM Methyltransferase Assays
Electrophoresis Clean-Up
Protease Assays
Concentration Systems
Phosphatase Assays
Contamination Removal
Peroxide Assay
Biotin Labeling
Carrier Proteins
Cell Surface Protein Labeling
Peptide Coupling Systems
Agarose Coupling Kits
Antibody Purification Resins
Fluorescent Dye Labeling Kits
Antibody Fragmentation Kits
Enzyme Labeling Systems
Coated Plates
Blocking Buffers Homobifunctional
Wash Buffers Heterobifunctional
Secondary Antibodies Optimizer Systems
Detection Reagents Cross-Linking Systems
Antibody Labeling Systems
Apoptosis Assays
DNA Isolation
Cytotoxicity Assays
Transformation & Screening
SAM Methyltransferase Assays
Polymerase Chain Reaction
Protease Assays
Agarose Electrophoresis
Phosphatase Assays
RNA Isolation
Peroxide Assay
Yeast Transformation
ELISA
Table Of Contents
Genomic DNA Purification 2 PCR 14
Introduction............................................................................2 DNA Polymerases...................................................................14
Purest DNA with OmniPrep. ................................................2 Taq Polymerase........................................................................14
OmniPrep . ..............................................................................2
Taq Polymerase (2X MasterMix).............................................14
OmniPrep for Tissue..............................................................2 Pfu Polymerase........................................................................14
OmniPrep for Blood................................................................3 Pfu Polymerase (2X MasterMix)..............................................14
OmniPrep for Plant.................................................................3 Deoxynucleotides...................................................................15
OmniPrep for Gram Positive Bacteria...................................3 Deoxynucleotides (dNTPs) Set................................................15
OmniPrep for Fungi................................................................3 Deoxynucleotides (dNTPs) Mix................................................15
OmniPrep for Yeast................................................................4 PCR Clean Up.........................................................................15
OmniPrep for Mouse Tail.......................................................4 Spin-OUT for PCR...................................................................15
OmniPrep Soil DNA................................................................4
Spin Format Isolation with GET...........................................4 RNA Purification 16
GET DNA Template.................................................................4
Tri-Xtract .................................................................................16
FEATURES
Isolate high quality genomic DNA
Suitable for fresh, frozen and fixed tissue Figure 5: XIT DNA from Blood scheme.
Yield 0.5-10g/mg tissue FEATURES
Suitable for archive quality DNA
Isolate high quality genomic DNA
Suitable for blood, bone marrow and buffy coat
Yield 10-15g/ml blood
Suitable for archive quality DNA
Figure 4: 10mg mouse liver was treated with the XIT Genomic DNA
from Tissue kit. The precipitated DNA was hydrated in 30l TE buffer and
3l was loaded onto an agarose gel.
Figure 6: Genomic DNA was isolated from 500l bloodusing the XIT
Cat. No. Description Size Genomic DNA from Blood Kit.
786-345 XIT DNA from Tissue For 250mg tissue
786-346 XIT DNA from Tissue For 2.5g tissue Cat. No. Description Size
786-347 XIT DNA from Tissue For 10g tissue 786-294 XIT DNA from Blood For 12.5ml blood
786-295 XIT DNA from Blood For 125ml blood
786-296 XIT DNA from Blood For 250ml blood
FEATURES
Figure 9: XIT DNA from FFPE Tissue scheme. Yield 1-5g/mg tissue
Suitable for archive quality DNA
FEATURES
Suitable for fresh, frozen and fixed tissue
Yield 0.5-10g/mg tissue
Suitable for archive quality DNA Cat. No. Description Size
786-297 XIT DNA from Plant Tissue For 2.5g tissue
Cat. No. Description Size
786-298 XIT DNA from Plant Tissue For 25g tissue
786-290 XIT DNA from FFPE Tissue For 250mg tissue
FEATURES
Yield 15-25g/ml bacterial culture Figure 15: XIT DNA from Yeast scheme.
Suitable for archive quality DNA
Cat. No. Description Size
786-339 XIT DNA from Gram positive bacteria For 25ml culture
786-340 XIT DNA from Gram positive bacteria For 250ml culture
XIT DNA from Gram Negative Bacteria Figure 16: Genomic DNA was isolated from 1ml overnight yeast culture, using
XIT DNA from Yeast kit. The DNA was resuspended in 50l and 5l or 10l
Designed for the isolation of genomic DNA from overnight were resolved on an agarose gel.
cultures. The kit uses three main steps that are rapid & efficient
bacterial lysis, complete protein precipitation, followed by DNA FEATURES
precipitation. This yields high quality genomic DNA. Yield 1-6g/mg tissue
Two sizes of kit are available for processing a total of 25 and Suitable for archive quality DNA
250ml of bacterial culture. The protocol is designed to use 1ml Cat. No. Description Size
overnight culture, however the protocol can be easily adapted for 786-348 XIT DNA from Yeast For 25ml culture
larger culture sizes. The purified DNA has a A260/A280 ratio between 786-349 XIT DNA from Yeast For 250ml culture
1.7 and 1.9 and is up to 200kb in size. The yield is approximately
20-40g/ml culture.
XIT Mitochondrial DNA
The XIT Mitochondrial DNA kit combines our FOCUS
Mitochondria technology with our XIT DNA isolation technology in
a single kit. The kit uses our proprietary Subcell buffers to lyse cells
and animal tissue and remove cellular proteins and nuceli from
a highly enriched, intact and active mitochondrial fraction. The
mitochondria are then lyzed with the included lysis buffer.
The lysate is treated with our LongLife Proteinase K to degrade
Figure 14: XIT DNA from Gram Negative Bacteria scheme. the proteins, which are then precipitated and the mitochondrial DNA
is isolated with alcohol precipitation.
FEATURES The kits is designed for ~100 preps consisting of 20x106
Yield 20-40g/mg tissue mammalian cells/ prep. The kit protocol also contains protocols for
Suitable for archive quality DNA isolation of mitochondra from soft (i.e. brain & liver) and hard (i.e.
Cat. No. Description Size cardiac & skeletal muscle) tissue.
786-337 XIT DNA from Gram negative bacteria For 25ml culture FEATURES
786-338 XIT DNA from Gram negative bacteria For 250ml culture Isolate high quality mitochondrial DNA
Suitable for cultured cells, soft and hard animal tissue
Cat. No. Description Size
786-301 XIT Mitochondrial DNA 100 preps
FEATURES
For animal and plant samples
Compatible with PCR and other downstream applications
APPLICATIONS
For the rapid preparation of DNA template
Cat. No. Description Size
786-014 Rapid DNA Template Prep 50 preps
Figure 17: OmniTemplate produces high quality genomic DNA template.
786-015 Rapid DNA Template Prep 100 preps
5l (lanes 1, 3, 5) or 15l (lanes 2, 4, 6) PCR reactions were loaded
onto a 2% agarose gel. Template was purified from 104 (lanes 1-2), 106
(lanes 3-4) or 2.8x106 (lanes 5-6) NIH3T3 cells. Yeast Geno-DNA-Template
FEATURES
Extraction of high quality DNA from yeast
Single tube genomic DNA isolation
Compatible with PCR and other downstream applications This extraction kit isolates high quality genomic DNA from yeast.
Suitable for blood, cells, and animal tissue samples The kit is based on a two-step lysis of yeast cells, using LongLife
No toxic chemicals, no phenol, no hazardous waste Zymolyase and LongLife Proteinase K, followed by the removal
of proteins and other cellular impurities by precipitation and
APPLICATIONS
centrifugation. The clean, genomic DNA is collected by precipitation
Designed for the rapid preparation of DNA template for large and the high yield of extracted DNA has an average 100kb length and
throughput microanalysis has A260/280 1.8-2.0.
High recovery of ready-to-use DNA templates in a short period of For extracting DNA from 100 x 1.5ml yeast cultures.
time
Cat. No. Description Size
786-013 OmniTemplate >100 preps
FEATURES
Isolation of high molecular weight genomic DNA
For animal tissues, cultured cells, whole blood, bacterial and yeast
Uses patented genomic Tube-O-DIALYZER to minimize sample
manipulation
APPLICATIONS
For DNA libraries, PCR amplification, Southern blot analysis, pulse-
field electrophoresis, and other applications where >100kb DNA is
required
Cat. No. Description Size
786-146 MegaLong 25 preps
786-147 MegaLong 50 preps
FEATURES
Simple, 5 minute protocol, as easy as picking a colony
No overnight cultures required
APPLICATIONS
Rapid colony screening, using restriction digestion analysis
Cat. No. Description Size
786-026 Plasmid Screening ToothPick 300 preps
ToothPick-PCR
FEATURES
Spin column format
Enhanced DNA binding affinity Figure 27: Genomic Tube-O-DIALYZER scheme.
Protocol is 5-10 minutes
No phenol/chloroform extraction or alcohol precipitations FEATURES
APPLICATIONS Rapid removal of impurities, RNA, small fragment DNA, etc.
PCR clean-up 100% sample recovery
Purifying DNA fragments after in-vitro labeling reactions Single tube for dialysis and storage, prevents sample loss
Removal of restriction enzymes from plasmid DNA For 0.2-2.5ml sample
Cleaning any DNA preparations from interfering agents No contamination
APPLICATIONS
Cat. No. Description Size
For the clean-up of genomic (>100kb) DNA by dialysis
786-356 GET CLEAN DNA 25 preps
Patented, single tube dialysis device to ensure no sample loss
786-357 GET CLEAN DNA 50 preps
Cat. No. Description Size
Aqueous mixture of dATP, dCTP, dGTP and dTTP. The dNTP Mix is a
ready-to-use aqueous solution containing dATP, dCTP, dGTP and dTTP,
each at a final concentration of 10 or 25mM. The Mix reduces the
number of pipeting steps and the risk of errors.
Applications
Ready to use in PCR, long-PCR, RT-PCR, cDNA synthesis, primer
extention and DNA labeling.
Cat. No. Description Size
786-443 Deoxynucleotide Mix [10mm], 0.5ml 5mole
786-442 Deoxynucleotide Mix [10mm], 5 x 0.5ml 25mole
786-457 Deoxynucleotide Mix [25mM], 1ml 25mole
786-458 Deoxynucleotide Mix [25mM], 5ml 125mole
786-459 Deoxynucleotide Mix [25mM], 25ml 250mole
PCR Clean Up
RNasin
RNase Inhibitor
RNasin is a ribonuclease inhibitor extracted from human placenta
with a molecular weight 51kDa. It inhibits the activity of RNase by
specifically binding up to RNase with a non-covalent bond. RNasin,
free of RNase or Nickase, can maintain its activity at pH from 5 to 8,
and the highest one at pH7.8. Concentration is 20~40 units/l.
Cat. No. Description Size
786-815 RNasin [20-40U/l] 1,000 units
FEATURES
Extract DNA fragments from agarose gels
Rapid DNA isolation
DNA ready for downstream applications, including ligations
Suitable for 100-20,000bp fragments
Compatible with TAE and TBE buffer gels
Figure 35: The G-CAPSULE procedure APPLICATIONS
Isolation of DNA fragments from agarose gels
FEATURES Cleaning and removing of contaminants from DNA samples
Rapid electroelution of nucleic acids and proteins
Cat. No. Description Size
Sample recovered in a small volume (25-50l)
Recovery is as high as 90% 786-358 GET AGAROSE DNA 50 preps
786-359 GET AGAROSE DNA 100 preps
APPLICATIONS
Extraction of >20bp DNA and RNA or for >4kDa proteins
geneEXIT
ACCESSORIES
G-CAPSULE Weight: Prevents G-CAPSULE from floating during
Isolation of nucleic acids from agarose gels
electroelution
The kit is based on pinkRESIN, a high capacity, proprietary
binding resin matrix for nucleic acids. pinkRESIN has an enhanced
binding affinity for DNA and RNA and resuspends with ease, thus
eliminating loss and damage of nucleic acids, a frequent problem
with other binding resin matrices for DNA and RNA isolation.
pinkRESIN is a non-toxic matrix.
geneEXIT method involves release of nucleic acid fragments
from gel pieces followed by capture of nucleic acids with pinkRESIN,
washing, and elution of the clean nucleic acid fragments in a suitable
buffer. The geneEXIT method can be carried out with or without the
use of spin columns; however the use of spin columns simplifies the
Figure 36: The G-CAPSULE weight protocols.
Cited References FEATURES
Chatterjee, S et al (2012) Acta Biochim Biophys Sin. 259:68 Rapid nucleic acid isolation
Chatterjee, S et al (2012) Acta Biochim Biophys Sin. 44:259 DNA & RNA ready for further applications, including ligations
Chatterjee, S et al (2012) Acta Biochim Biophys Sin. 10:1093 Suitable for 100-20,000bp fragments
Cardi, D et al (2010) J Biol Chem. 285:26406
Crosslin, J (2009) HortScience. 44:1790 Compatible with TAE and TBE buffer gels
Li, X et al (2004) Euro J of Phycology. 39:73 Supplied with or without spin columns
Beeson, K et al (2002) Microbiology. 148:179
Yeager, M et al (2001) Circ Res. 88:2e
APPLICATIONS
Robu, M et al (2001) PNAS. 98:8211 Isolation of DNA/RNA fragments from agarose gels
Dudley, E et al (2001) Microbiology. 147:215 Cleaning contaminants from DNA & RNA samples
Brezinschek, Hans-Peter et al (2000) Int Immunol. 12:767
Tanaka, K et al (1999) Yeast. 15:1133 Cat. No. Description Size
Cat. No. Description Size 786-85 geneEXIT 100 preps
786-001 G-CAPSULE 55/box 786-86 geneEXIT with spin columns 100 preps
786-004 G-CAPSULE Weight 1
Bromophenol Blue
Ethidium Bromide
highly toxic ethidium bromide stain. High concentrations of ethidium
Xylene Cyanol
bromide are either added to the molten agarose prior to pouring or
to a large volume of staining buffer to stain the nucleic acids after
electrophoresis. Either method results in a large amount of ethidium
bromide to handle and dispose of.
SDS
Cat. No. Description
The Glow Dyes are designed to minimize the use of ethidium
DNA Glow Dyes [6X]
bromide and the risk associated with the regular use of ethidium
bromide. Glow Dyes have been specifically formulated for nucleic 786-103 Glow BromoBlue Dye Yes No No Yes
acid electrophoresis and contain an optimized concentration of 786-104 Glow CyanoBlue Dye Yes Yes No Yes
buffer agents, tracking dyes and ethidium bromide. These loading 786-105 Glow CleanAway Dye Yes Yes Yes Yes
dyes reduce exposure and many of the problems associated with DNA Universal Dyes [6X]
ethidium bromide use and disposal. Simply add an appropriate 786-100 BromoBlue Universal Dye Yes No No No
volume of Glow Dyes to your sample, load the gel and then visualize 786-101 CyanoBlue Universal Dye Yes Yes No No
with UV; no need for additional ethidium bromide staining or addition 786-102 CleanAway Universal Dye Yes Yes Yes No
of ethidium bromide to the agarose. RNA Glow Dyes [2X]
The DNA Glow Dyes are Ficoll based and are supplied at a 6X 786-107 Glow RNA Dye Yes Yes No Yes
concentration and the RNA Glow Dyes are formamide based and are
RNA Universal Dyes [2X]
at a 2X concentration.
786-106 Universal RNA Dye Yes Yes No No
The DNA and RNA Loading Dyes are offered in multiple formats to
meet all your needs and have variations in the following components:
Dyes: The loading dyes use Bromophenol Blue that migrates at
~300bp (0.7-1.7% agarose) or ~100bp (2.5-3.0% agarose) to
highlight the migration front and Xylene Cyanol FF (CyanoBlue
Loading Dyes) that migrates at ~4,000bp (0.7-1.7% agarose)
or ~800bp (2.5-3.0% agarose) to help resolve higher molecular
weight nucleic acids.
SDS: The detergent SDS (sodium dodecyl sulfate) is supplied in the
CleanAway Loading Dyes and these are recommended for use
with DNA that has a high level of DNA binding proteins. The SDS
eliminates DNA-protein interactions, which prevents poor DNA
migration, DNA sticking to the wells or band shifts. In addition,
SDS treatment prevents long cohesive ends reannealing and Figure 38: DNA/RNA loading dyes
producing artifactual DNA bands.
Ethidium Bromide: The benefits of ethidium bromide in the Glow
Loading Dyes is highlighted above. For researchers who want the
benefits of our loading dyes without the ethidium bromide, try our
Universal Loading Dyes.
FEATURES
Intense DNA/RNA bands with little background or band distortion
Compatible with all agarose or acrylamide gel electrophoresis
Glow RNA Dye is ideal for quick screenings of RNA preps
There is no need to use toxic formaldehyde agarose gels for
checking RNA integrity with Glow RNA Dye
Also available: Universal Loading Dyes containing different tracking
dyes, no ethidium bromide, and the option of added SDS
APPLICATIONS
Suitable as sample loading dye for electrophoresis application
Tracking electrophoresis migration
Visualization of nucleic acids
GELPLATE-clean
LabSafe Nucleic Acid Stain
Spray and wipe; specifically formulated for
LabSafe Nucleic Acid Stain is a new and safe nucleic acid electrophoresis gel plates
stain for the visualization of double-stranded DNA, single-stranded
DNA, and RNA in agarose gels. The dyes are developed to replace Clean your electrophoresis plates with GELPLATE-clean and
toxic Ethidium Bromide (a potent mutagen), commonly used in gel prevent poor electrophoretic band migration, band distortions,
electrophoresis for visualization of nucleic acids in agarose gels. and poor image development. Simply spray and wipe clean your
LabSafe Nucleic Acid Stain is non-carcinogenic by the Ames-test. electrophoresis plates with GELPLATE-clean.
The results are negative in both the mouse marrow chromophilous FEATURES
erythrocyte micronucleus and mouse primary spermatocyte Suitable for sequencing and protein electrophoresis gel plates
chromosomal aberration tests. Removes proteins, nucleic acids, fats, lipids, radioisotopes and
LabSafe Nucleic Acid Stain emits green fluorescence when other contaminants from electrophoresis gel plates
bound to dsDNA and red fluorescence when bound to ssDNA or RNA. Supplied in 2 x 250ml easy to apply spray bottles
It has two excitation wavelength peaks when bound to nucleic acid, Refill solutions are offered separately
at 290nm and 490nm. APPLICATIONS
Cleans sequencing and electrophoresis gel plates
Cat. No. Description Size
786-140 GELPLATE-clean Spray 2 x 250ml
786-140RF GELPLATE-clean Refill 2 x 1L
Figure 39: Plasmid DNA stained with LabSafe Nucleic Acid Stain 786-140RF-I GELPLATE-clean Refill for Solution I 1L
786-140RF-II GELPLATE-clean Refill for Solution II 1L
Cat. No. Description Size
786-409 LabSafe Nucleic Acid Stain [10,000X] 1ml
CITED REFERENCES
Guo, H. et al (2010) Antimicrob. Agents Chemother. doi:10.1128/AAC.00989
Guo, H. et al (2007) J. Virology 81: 10072
Guo, H. et al (2007) J. Virology 81: 12472
Dougherty, A. M. et al (2007) Antimicrob. Agents Chemother. 51:4427
Hodson, J. et al (2003). Nuc. Acid Res. e31: e134
Kim, Mee-Jung et al (2002) PNAS. 99: 10096
Moraleda, G. and Taylor, J. (2001) J. Virol. 75: 10161
HYB-LINK
Formamide-free hybridization buffer
Hyb-LINK is a high performance formamide free hybridization
solution for Northern, Southern and Dot Blots, which increases
sensitivity of random-primer DNA probes. Hyb-LINK capitalizes
on increased sensitivity of blot hybridization without increasing
background and therefore provides reliable and accurate
hybridization.
Hyb-LINK is compatible with both RNA and DNA probes.
Cat. No. Description Size
786-405 Hyb-LINK 125ml
786-406 Hyb-LINK 4 x 125ml
FEATURES
For extraction of genomic DNA from yeast cultures
Compatible with PCR and other downstream applications
APPLICATIONS
Designed for the preparation of high yield, genomic DNA template
from yeast
Cat. No. Description Size
786-134 Yeast Geno-DNA-Template 100 preps
LongLife Zymolyase
Figure 43: LongLife Enzymes are highly stable. Each enzyme preparation
A high performance LongLife Zymolyase preparation is supplied was tested over a period of 4 weeks at 37C & compared with LongLife
in a ready-to-use solution form. The enzyme contains high yeast lytic enzyme preparations stored at -20C. LongLifeZymolyase (1.5units/l)
activity with low non-specific activity. Suitable for yeast cell lysis, was tested with freshly grown yeast suspension by monitoring the decrease in
spheroplast preparation, glucan hydrolysis and more. The enzyme absorbance of the suspension. LongLife Lysozyme (1500units/l) was tested
preparation has been stabilized and can be stored at -20C. by monitoring the decrease in the absorbance of Micrococcus lysodeikticus
suspension. LongLife Proteinase K (5mg/ml) was assayed with our Protease
Assay kit (Cat. No. 82023-260). No measurable loss of activity was noticed.
Cited References
LongLife RNase
Tashiro, R. et al (2010) Crop Sci. 50:1260
LongLife Lysozyme
Butcher, B.G. et al (2011) J Bacteriol. 193:4598
Ermolova, N. et al (2011) Hum.Mol Genet. 20:3331
Markel, E. et al (2011) J Bacteriol. 193:5775
LongLife Proteinase K
Figure 42: LongLife Zymolyase are highly stable. The enzyme Whitaker, V. M. et al (2007) J Amer Soc.Hort Sci.132:534
preparation was tested over a period of 4 weeks at 37C: and compared
with LongLife enzyme preparations stored at -20C. LongLife Cat. No. Description Size
Zymolyase (1.5units/l) was tested with freshly grown yeast suspension 786-036 LongLife Zymolyase [1.5U/l] 2 x 0.5ml
by monitoring the decrease in absorbance of the suspension. 786-037 LongLife Lysozyme [1,500U/l] 2 x 0.5ml
786-042 LongLife PE LB Lysozyme [1,500U/l] 2 x 0.5ml
CITED REFERENCES
Gray, P. et al (2006) Mol Cell Prot. 6:514
786-038 LongLife Proteinase K [5mg/ml] 2 x 0.5ml
Saribas, A. et al (2004) Glycobiology 14:1217 786-039 LongLife Nuclease [10U/l] 2 x 0.5ml
786-040 LongLife RNase [10U/l] 2 x 0.5ml
Cat. No. Description Size
786-041 LongLife DNase 0.5ml
786-036 LongLife Zymolyase 2 x 0.5ml
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Updated: February 11, 2014 www.GBiosciences.com