Cloning

<h2 data-section-id="k59sk3" data-start="0" data-end="52">What are transformation kits and competent cells?</h2>
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Transformation kits and competent cells consist of E. coli strains prepared to take up exogenous plasmid DNA, either through chemical competence (calcium chloride treatment, transformed via heat shock) or electrocompetence (transformed via electroporation), along with associated transformation buffers and protocols. Related entities include transformation efficiency (transformants per microgram), heat-shock and electroporation methods, strain genotype (recA, endA deficiency for plasmid stability), and downstream plasmid recovery for sequencing or transfection.
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<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li>Mix &amp; Go! Transformation Kits, rapid protocols for preparation of competent E. coli in&nbsp;</li>
<li>ProClone&trade; Competent Cells with optimized E. coli strains for cloning and library generation.</li>
<li>Specialized Competent E. coli Strains ready-made JM109, DH5 Alpha, and Zymo 10B cells for regular and complex assembly.</li>
<li>Autolysis Competent Cells: designed for simplified cell lysis and streamlined protein expression using XJa and XJb strains</li>
<li>Frozen-EZ Yeast Transformation Kits for genetic manipulation of yeast strains.</li>
</ul>
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<h2 data-section-id="1ld88i5" data-start="812" data-end="876">How to choose transformation kits and competent cell products</h2>
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Chemically competent vs. electrocompetent cells
Chemically competent cells use a heat-shock protocol and are generally easier to handle for routine cloning, while electrocompetent cells require an electroporator but can achieve higher transformation efficiency for large plasmids, low-copy constructs, or DNA with modifications that reduce chemical transformation efficiency.
General cloning strains vs. protein expression strains
General cloning strains are selected for high transformation efficiency and genetic features (such as recA and endA deficiency) that improve plasmid stability and quality for downstream sequencing, while protein expression strains carry features supporting recombinant protein production, such as T7 RNA polymerase expression for IPTG-inducible systems; using a cloning strain for protein expression (or vice versa) generally gives suboptimal results.
Transformation efficiency requirements
Routine cloning of standard-sized plasmids works well with competent cells rated at 10^6&ndash;10^8 transformants per microgram, while ligation reactions with very low DNA concentrations (such as multi-fragment seamless assembly products) may benefit from higher-efficiency cells rated above 10^8 transformants per microgram.
Transformation kits may include a positive control plasmid and competent cells for validating transformation efficiency before using precious experimental DNA; this is particularly useful when troubleshooting low transformation yields to distinguish competent cell quality issues from ligation or assembly reaction issues.
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<h2 data-section-id="1fljvas" data-start="3947" data-end="3972">Specifications context</h2>
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Competent cell transformation efficiency is reported in transformants per microgram of standard control plasmid DNA, with general cloning strains commonly rated at 10^6&ndash;10^8 and high-efficiency formulations exceeding 10^8 transformants per microgram. Cells are typically supplied as frozen aliquots (e.g., 50&ndash;100 microliters per tube) for single-use transformations, shipped on dry ice and stored at -80&deg;C, with repeated freeze-thaw substantially reducing efficiency.&nbsp;
This sub-category is part of MBP's cloning catalog alongside ligation independent cloning kits and chemicals/reagents, the upstream steps of a cloning workflow. Labs producing custom lentivirus for immortalization can also review MBP's cell immortalization catalog. MBP's specialist team can help confirm strain genotype and transformation efficiency requirements for a specific cloning or expression project before order placement.
<a href="https://mbpinc.net/contact-us">Contact the expert team</a> at MBP today and get high-quality transformation kits and competent cells for your lab.

Transformation Kits & Competetent Cells

MSDS

DataSheet

Protocol

Mix & Go! Competent Cells - XJa(DE3) Autolysis (10 x 100 µl)

T3031 Mix &amp; Go! Competent Cells — XJa(DE3) Autolysis by Zymo Research provides 10 × 100 µl aliquots of E. coli with 10⁸–9⁹ CFU/µg transformation efficiency and L-Arabinose-triggered λ endolysin autolysis for simplified protein expression and purification. Includes 1 ml 500× L-Arabinose; stored at -70°C to -80°C. MBP is a registered vendor for Howard Hughes Medical Institute, Vanderbilt University, MD Anderson Cancer Center, and many more leading research institutes, serving labs across the United States, Canada, and internationally.

Autolysis

DNA Extraction

DNA Stability

Genotype

Processing Time

Product Storage

Protein Expression

Manufacturer

Shipping Condition

Storage Condition

Mix & Go! Competent Cells - XJb Autolysis (10 x 100 µl)

T3041 by Zymo Research is the Mix &amp; Go! XJb Autolysis Competent Cells — an E. coli expression strain achieving 108–109 transformants/µg and 80–90% autolytic lysis within 10 minutes via L-Arabinose induction. Lon-/OmpT- for improved protein yields. Supplied as 10 × 100 µl with 1 ml 500× L-Arabinose. Store at -70°C to -80°C.

Cell Growth

Transformation Efficiency

Mix & Go! Competent Cells - XJb(DE3) Autolysis (10 x 100 µl)

Mix &amp; Go! Competent Cells - XJb(DE3) Autolysis (T3051) by Zymo Research — 10 × 100 µl. 108–109 transformants/µg; no heat shock. L-arabinose-induced autolysis: 80–90% cell lysis in ~10 min by freeze-thaw. DE3 lysogen for T7 expression. Lon-/OmpT- protease-deficient. Includes 500× L-arabinose. Storage −70°C to −80°C.

Mix & Go! Competent Cells - Zymo 10B

Mix &amp; Go! Competent Cells - Zymo 10B by Zymo Research — &gt;108 transformants/µg pUC19 without heat shock. Available as T3019 (10 × 100 µl) or T3020 (96 × 50 µl, 12 × 8-Tube Strips). Storage &lt;−70°C; 6-month integrity guarantee. Genotype: F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara leu) 7697 galU galK rpsL nupG λ-.

Additional Info

Mix & Go! Competent Cells - Zymo 10B (10 x 100 µl)

Mix & Go! Competent Cells - Zymo 10B (96 x 50 µl)

<h2 data-section-id="v1drft" data-start="0" data-end="43">What are cloning chemicals and reagents?</h2>
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Cloning chemicals and reagents cover the enzymes, buffers, and molecular biology-grade chemicals used in DNA fragment preparation, modification, and purification within a cloning workflow, including restriction endonucleases, T4 DNA ligase and other DNA-modifying enzymes (phosphatases, polymerases), reaction buffers, and plasmid extraction reagents. Related entities include restriction enzyme digestion, DNA dephosphorylation/phosphorylation, plasmid DNA purification (column-based and magnetic bead-based), and enzyme unit definitions.
&nbsp;
<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" role="presentation">Targeted XJa and XJb strains for faster recovery of proteins.</li>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" role="presentation">Mix &amp; Go! solutions for optimized and dependable bacterial transformation.</li>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" role="presentation">Culture Media &amp; Supplements used for yeast research, and include YPD Plus and mating pheromones.</li>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" role="presentation">Extraction and Lysis Reagents include reliable genomic preparation and yeast protein kits.</li>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" role="presentation">Chromogenic Substrates and Enzymatic Inducers with highly pure X-GAL and IPTG for cloning and blue-white screening.</li>
</ul>
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<h2 data-section-id="1daajgc" data-start="866" data-end="913">How to choose cloning chemicals and reagents</h2>
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Restriction enzyme selection and buffer compatibility
Restriction enzymes require specific reaction buffers, and many modern enzyme lines use a single universal buffer compatible across multiple enzymes to simplify double-digest reactions; confirm buffer compatibility when combining enzymes from different suppliers or using older single-buffer-per-enzyme formulations.
Enzyme unit definitions and reaction scale
Restriction enzyme and ligase activity is defined in units per microliter, with one unit typically defined as the amount needed to digest or ligate a defined quantity of standard substrate DNA under specified conditions in a set time; scale enzyme quantity to the amount of DNA being processed rather than assuming a fixed volume works for all reaction sizes.
Plasmid purification grade
Plasmid DNA extraction reagents range from molecular biology grade, suitable for general cloning and sequencing, to transfection-grade or endotoxin-free formulations required for applications such as lentiviral packaging or mammalian cell transfection, where endotoxin contamination can reduce transfection efficiency or cause cell stress.
Dephosphorylation for preventing vector self-ligation
Alkaline phosphatase treatment of a linearized vector before ligation removes 5' phosphate groups, preventing vector self-ligation and improving the ratio of recombinant clones; this step is particularly useful when the vector and insert share compatible sticky ends that could otherwise religate without insert incorporation.
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<h2 data-section-id="1fljvas" data-start="3459" data-end="3484">Specifications context</h2>
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Restriction enzymes and ligases are supplied at defined unit concentrations (commonly 10&ndash;20 units per microliter for restriction enzymes), with reaction buffers formulated for specific pH and salt conditions optimized for enzyme activity. Plasmid purification kits are differentiated by yield (micrograms of DNA recovered), purity (A260/A280 ratio), and endotoxin level (endotoxin units per microgram), with endotoxin-free kits specifying levels below a defined threshold suitable for transfection applications.&nbsp;
This sub-category is part of MBP's cloning catalog alongside ligation independent cloning kits and transformation kits and competent cells, used downstream of enzyme-based fragment preparation. Labs producing custom lentivirus for immortalization can also review MBP's cell immortalization catalog. MBP's specialist team can help confirm enzyme buffer compatibility and plasmid purification grade for a specific downstream application before order placement.
<a href="https://mbpinc.net/contact-us">Contact the expert team </a>at MBP today to request a quote for high-purity chemicals and reagents.

Chemicals/Reagents

Mix & Go! Dilution Buffer

Mix & Go! Dilution Buffer by Zymo Research is used to dilute 2X concentrated buffers to 1X working concentration in the Mix & Go! E. coli Transformation System (T3001/T3002 kits). Available in 20 ml and 60 ml formats, stored at 0–8°C, it ensures optimal conditions for high-efficiency chemical transformation of E. coli.

Mix & Go! Dilution Buffer (20 ml)

Mix & Go! Dilution Buffer (60 ml)

Mix & Go! E. coli Transformation Kit (up to 20 ml)

Mix &amp; Go! E. coli Transformation Kit T3001 by Zymo Research enables preparation of up to 20 mL of competent cells with 10⁸–10⁹ transformants/µg pUC19 and no heat shock required. Compatible with DH5α, BL21, JM109, TG1, and XL10 Gold. Competent cell preparation completes in ~1–2 hours.

Compatible Strains

Mix & Go! E.coli Transformation Buffer Set (up to 60 ml)

<h2 data-section-id="b4wzzk" data-start="0" data-end="47">What is bacterial culture media for cloning?</h2>
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Bacterial culture media for cloning workflows includes LB (Luria-Bertani) broth and agar, the standard general-purpose media for E. coli growth, SOC media used for post-transformation recovery, and Terrific Broth for higher-density culture in plasmid preparation. Pre-made agar plates with antibiotic selection (ampicillin, kanamycin, chloramphenicol, and others matching common selection markers) are used to isolate colonies carrying the desired plasmid. Related entities include antibiotic selection markers matching transformation kit resistance genes, post-transformation recovery time, and plasmid yield scaling with culture volume and media richness.
&nbsp;
<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li dir="ltr" role="presentation">ZymoBroth Growth Media for efficient transformation workflows to generate highly competent bacterial cells</li>
<li dir="ltr" role="presentation">Overexpression Broth (OB) optimized for increasing recombinant protein yields with high-density nutrient media.</li>
<li dir="ltr" role="presentation">Expansion Broth (EB) for the faster and more reliable expansion of microbial cultures.</li>
<li dir="ltr" role="presentation">Dual Media Sets for integrated growth and induction workflows with the help of combined EB and OB systems</li>
</ul>
&nbsp;
<h2 data-section-id="9xi4x5" data-start="919" data-end="971">How to choose bacterial culture media for cloning</h2>
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LB broth/agar vs. Terrific Broth
LB broth and agar are the standard general-purpose media for routine cloning, colony selection, and small-scale plasmid preparation, while Terrific Broth provides higher nutrient density supporting higher bacterial cell density, making it useful when maximizing plasmid yield in larger-scale preparations.
SOC media for post-transformation recovery
SOC media is richer than standard LB and supplemented with glucose and magnesium. It is commonly used for a brief recovery incubation immediately after heat-shock or electroporation transformation, helping transformed cells recover before plating on selective agar. Some transformation protocols specifically recommend SOC rather than LB for this recovery step.
Antibiotic selection matching
Pre-made antibiotic-selective agar plates must match the resistance marker carried by the transformed plasmid, whether ampicillin, kanamycin, chloramphenicol, or another selection marker. Using the wrong antibiotic&mdash;or no antibiotic at all&mdash;will not properly select for plasmid-containing colonies and may allow unwanted bacterial growth.
Liquid vs. agar plate formats and shelf life
Liquid media (broth) is used for post-transformation recovery and culture scale-up, while agar plates are used for colony isolation and selection. Pre-made antibiotic plates have a defined shelf life because antibiotics gradually lose activity over time, making expiration dates particularly important for antibiotics with lower stability.
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<h2 data-section-id="1fljvas" data-start="3523" data-end="3548">Specifications context</h2>
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LB broth and agar are formulated with defined concentrations of tryptone, yeast extract, and sodium chloride, with agar plates commonly supplied in standard Petri dish sizes (for example, 100 mm) and containing antibiotic concentrations matched to common resistance markers, such as 100 &micro;g/mL ampicillin or 50 &micro;g/mL kanamycin. SOC recovery is typically performed as a short incubation, often around one hour at 37&deg;C with shaking, before plating cells on selective agar.&nbsp;
This sub-category is part of MBP's cloning catalog alongside transformation kits and competent cells, the immediately preceding step, and chemicals/reagents for enzyme-based fragment preparation. Labs scaling up plasmid production for lentivirus packaging in immortalization workflows can also review MBP's cell immortalization catalog. MBP's specialist team can help confirm antibiotic concentration and media format requirements for a specific selection marker before order placement.
<a href="https://mbpinc.net/contact-us">Contact the expert team</a> at MBP today to request a quote for high-quality culture media.

Culture Media

Overexpression Broth (OB)

Overexpression Broth OB — M3013-100 by Zymo Research — is a 100 mL specialized bacterial medium for high-level recombinant protein expression, designed for use alongside Expansion Broth (EB) in a two-phase culture protocol. It stores at room temperature and is also available in a 500 mL format (M3013-500). Order from MBP via <a href="https://www.mbpinc.net/quickorder">Quick Order</a> or contact <a href="mailto:customerservice@mbpinc.net">customerservice@mbpinc.net</a> for volume pricing.

Overexpression Broth™ (OB™) (100 ml)

Overexpression Broth™ (OB™) (500 ml)

Rattler Plating Beads

Rattler Plating Beads by Zymo Research — 4.5 mm solid glass beads for spreading bacteria and yeast on agar plates without a spreader or flame. Reusable after bleach wash and autoclaving. Available as S1001 (1×230 g, sterile), S1001-5 (5×230 g, sterile), and S1001-B (25 kg bulk bag, non-sterile).

Material

Packaging

Rattler™ Plating Beads (230 g Bottle)

Rattler™ Plating Beads (5 x 230 g Bottles)

Rattler™ Plating Beads, Bulk (25 kg Bag)

X-GAL (5-bromo-4-chloro-3-indolyl β-D-galactopyranoside)

X1001-25 by Zymo Research is X-GAL, a ready-to-use 2% w/v sterile solution in DMF for blue-white colony screening and β-galactosidase detection. Available as X1001-25 (5 × 5 ml) and X1001-5 (1 × 5 ml). Formulated to minimize false positives. Store at -20°C. Made in the USA by Zymo Research.

Concentration

5-bromo-4-chloro-3-indolyl β-D-galactopyranoside (X-GAL) (5 x 5 ml)

5-bromo-4-chloro-3-indolyl β-D-galactopyranoside (X-GAL) (5 ml)

XJa (DE3) Autolysis Glycerol Stock (1 Tube)

XJa (DE3) Autolysis Glycerol Stock T5031 by Zymo Research — 1 mL glycerol stock with 500× L-arabinose — provides 80–90% cell lysis after a single freeze-thaw cycle (~10 min post-harvest). The DE3 lysogen supports T7 promoter–based recombinant protein expression; growth requires 1 mM Mg²⁺ supplementation. Storage at –70°C to –80°C. Order via <a href="https://www.mbpinc.net/quickorder">Quick Order</a> or contact <a href="mailto:customerservice@mbpinc.net">customerservice@mbpinc.net</a>.

XJa Autolysis Glycerol Stock (1 Tube)

T5021 by Zymo Research is the XJa Autolysis Glycerol Stock — an EndA-/RecA- E. coli expression strain with L-Arabinose-inducible autolysis achieving 80–90% cell lysis in 10 minutes. Supplied as 1 tube with 1 ml 500× L-Arabinose. Handles plasmids up to 10 kb. Store at -70°C to -80°C. For protein expression and high-quality nucleic acid extraction.

XJb (DE3) Autolysis Glycerol Stock (1 Tube)

T5051 by Zymo Research is the XJb (DE3) Autolysis Glycerol Stock (1 ml, 1 tube), a self-lysing E. coli expression strain with DE3 T7 RNA polymerase, Lon⁻ OmpT⁻ background, and arabinose-inducible λ endolysin for 80–90% cell lysis within 10 minutes. Eliminates sonication in recombinant protein workflows. Supplied with 500× L-Arabinose. Store at -70°C to -80°C.