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Plasmid Miniprep/Midiprep Kit
The ZY PurePrep Plasmid Miniprep/Midiprep Kit combines SDS alkaline lysis with silica-membrane adsorption to rapidly isolate high-purity plasmid DNA from E. coli cultures.
The purified DNA is suitable for routine molecular biology applications including restriction digestion, sequencing, PCR, ligation, and transformation.
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  • 2-4mL

    Culture volume for miniprep

  • 4-6mL

    Culture volume for midiprep

  • ~70μg

    Midiprep column capacity

  • 1.5years

    Stable storage at room temperature

首页 > Products > Molecular Biology Reagents > Plasmid Miniprep/Midiprep Kit >
Product Overview
Ordering Information
Product Highlights
Product Specifications
Procedure
Storage and Shipping
Key Operating and Quality Points
Applications
Frequently Asked Questions
Product Overview
Small-scale preparation
Medium-scale preparation

ZY PurePrep Plasmid Small-scale/Medium-scale Preparation Kit

Efficient extraction, flexible adaptation

Using the classic SDS-alkaline lysis method combined with silica membrane adsorption technology, high-purity plasmid DNA can be rapidly and efficiently extracted from E. coli cultures.

The extracted plasmid DNA can be directly used for routine molecular biology experiments such as restriction digestion, sequencing, PCR, ligation, and transformation.

  • Mature process with consistent results between batches
  • No need for phenol/chloroform extraction and ethanol precipitation
  • Flexible adaptation for small and medium-scale preparations
Plasmid Miniprep/Midiprep Kit

Small-scale plasmid preparation

Suitable for daily clone screening and routine validation

The small-scale kit is suitable for 2–4 mL overnight cultures and can be extended to 4–6 mL, making it suitable for the rapid identification of positive clones and meeting the needs of routine screening such as sequencing and restriction digestion verification.

  • Item Number: ZY-PP-MINI-200
  • Specification: 200 times
  • Elution Volume: 50–100 μL
  • Operation Time: Approximately 25–35 min
Small-scale plasmid preparation

Medium-scale plasmid preparation

Supports downstream experiments with multiple tubes and batches

Medium-scale kits are suitable for 4–6 mL overnight bacterial culture; for low-copy plasmids, it can be extended to 10 mL to obtain a larger amount of high-purity plasmid.

  • Item Number: ZY-PP-MED-50
  • Specification: 50 times
  • Binding Column Capacity: Approximately 70 μg
  • Typical yield: approximately 30–50 μg (4 mL LB, high copy)
  • Operation time: approximately 35–45 min
Medium-scale plasmid preparation

Ordering Information

Two specifications: small and medium, flexibly suitable for different experiment scales
Product Name Catalog No. Size Applicable scenarios Price
ZY PurePrep
Plasmid Small-Scale Preparation Kit		 ZY-PP-MINI-200 200 times Recombinant clone primary screening, restriction digestion identification,
Pre-sequencing preparation, PCR template preparation
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ZY PurePrep
Plasmid mid-prep kit		 ZY-PP-MED-50 50 times Large-scale transformation, multi-tube restriction digestion, in vitro transcription,
Cell transfection, plasmid strain preservation
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Product Highlights

Stable and reliable, high purity for direct use, easy operation, flexible specifications, economical and efficient
  • Classic technology · Stable and reliable
    SDS-alkaline lysis combined with silica membrane adsorption, mature process, consistent results between batches.
  • High purity · Direct use downstream
    Effectively removes impurities such as proteins, RNA, and gDNA, meets A260/A280 standards, can be directly used in various downstream experiments.
  • Easy to operate · No toxic reagents
    Centrifugal column method, no phenol/chloroform extraction, no ethanol precipitation, safe and time-saving.
  • Flexible specifications · Suitable for different amounts of bacteria
    Small scale suitable for 2–4 mL and can be extended to 4–6 mL; medium scale suitable for 4–6 mL, low copy can be up to 10 mL.
  • Economical and efficient · Daily preferred choice
    High cost performance, suitable for high-throughput routine experiments such as clone screening, restriction digestion verification, and sequencing.

Product Specifications

Specifications, performance parameters, and complete kit components
Parameters Small-scale preparation kit Medium-scale preparation kit
Catalog No. ZY-PP-MINI-200 ZY-PP-MED-50
Size 200 times 50 times
Applicable bacterial culture volume 2–4 mL (expandable to 4–6 mL) 4–6 mL (low copy can reach 10 mL)
Column binding capacity — About 70 μg
Typical yield μg to tens of μg (depending on copy number and bacterial culture amount) About 30–50 μg (4 mL LB, high copy)
Elution volume 50–100 μL 100 μL
Operating Time Approximately 25–35 min Approximately 35–45 min
Storage Conditions 15–25℃ for 1.5 years; 2–8℃ for long-term 15–25℃ for 1.5 years; 2–8℃ for long-term
Transport Conditions Transport at room temperature Transport at room temperature (use ice packs if >30℃)

Note:Typical yields are for reference only. Actual yields depend on plasmid copy number, strain status, culture conditions, and other factors.

Procedure

Bacterial collection, lysis and neutralization, binding and washing, elution and detection
  • Collection and Resuspension
    Centrifuge to collect overnight cultured bacteria and fully resuspend in Solution I containing RNase A.
  • Lysis and Neutralization
    Add Solution II and gently mix to lyse, then add Solution III to neutralize and centrifuge to clarify.
  • Binding and Washing
    Transfer the supernatant to the adsorption column to allow the plasmid to bind to the silica membrane, and complete the washing according to specifications.
  • Elution and Detection
    Thoroughly dry the adsorption column, add Elution Buffer for elution, and use after electrophoresis detection or store at -20°C.

Small-scale Preparation (ZY-PP-MINI)

Detailed operational conditions for 2–4 mL overnight bacterial culture
Steps Operational Conditions
Column Equilibration Add 200 μL of Buffer CBS, centrifuge at 12,000 rpm for 1 min, discard the supernatant
Collect the bacterial cells Centrifuge 2–4 mL of bacterial culture at 8,000 rpm for 1 min; for 4–6 mL, combine in 2–3 portions
Resuspend Add 250 μL of Solution I (with RNase A added), fully suspend
Lysis Add 250 μL of Solution II, gently invert 6–8 times, room temperature no more than 5 min
Neutralize Add 350 μL of Solution III, invert 8–10 times, let stand for 2–5 min, centrifuge at 12,000 rpm for 5–10 min
Binding Transfer the supernatant to the adsorption column, centrifuge at 12,000 rpm for 1 min
Washing 500 μL of Wash Solution, centrifuge at 12,000 rpm for 1 min, repeat once
Drying Empty column, centrifuge at 12,000 rpm for 2 min, leave the lid open at room temperature for a few minutes
Elution 50–100 μL Elution Buffer, place at 37°C for 2 min, centrifuge at 12,000 rpm for 1–2 min
Detection Take 1–3 μL for electrophoresis detection, use immediately or store at -20°C

Medium-scale preparation (ZY-PP-MED)

Detailed operating conditions for 4–6 mL overnight bacterial culture
Steps Operational Conditions
Column Equilibration Add 200 μL of Buffer CBS, centrifuge at 12,000 rpm for 1 min, discard the supernatant
Collect the bacterial cells 4–6 mL bacterial culture, centrifuge at 8,000 rpm for 1 min; for low-copy plasmids, 10 mL can be used
Resuspend Add 500 μL of Solution I (with RNase A added), fully resuspend
Lysis Add 500 μL of Solution II, gently invert 4–6 times, leave at room temperature for no more than 5 minutes
Neutralize Add 700 μL of Solution III, invert 8–10 times until cloudy white, stand for 2–5 minutes, centrifuge at 12,000 rpm for 5–10 minutes
Binding Transfer the supernatant to the spin column in two portions, centrifuge at 6,000 rpm at room temperature for 1 minute
W1 wash Add 500 μL of W1 Solution, centrifuge at 12,000 rpm for 1 minute
Washing Add 500 μL of Wash Solution, centrifuge at 12,000 rpm for 1 minute, repeat once
Drying Centrifuge empty column at 12,000 rpm for 1 min, open the lid and leave at room temperature for a few minutes
Elution Add 100 μL Elution Buffer, incubate at 37°C for 2 min, centrifuge at 12,000 rpm for 1 min
Detection Take 1–3 μL for electrophoresis detection, use immediately or store at -20°C

Storage and Shipping

Store reagents according to conditions to ensure long-term stability and good extraction results
  • Stable at room temperature
    The remaining components can be stably stored under dry conditions at 15–25℃ for 1.5 years; the solution portion has a shelf life of 18 months.
  • Long-term low-temperature storage
    For long-term storage or when ambient temperature is too high, please store at 2–8℃.
  • Transport Conditions
    Room temperature transport; in summer, when the temperature exceeds 30℃, ice packs are recommended.
  • Solution usage recommendations
    Solution II / Solution III can be stored at room temperature; pre-cooling Solution III at 4℃ before experiments can achieve better results.
  • Usage statement
    For research use only, not for clinical diagnosis or treatment.

Key Operating and Quality Points

Strictly control resuspension, lysis, washing, and elution conditions to ensure plasmid yield and purity.
Control stepsDocument requirementsPurpose
RNase A preparationAdd all to Solution I, mix well, store at 4°C, and label the date; for long-term storage, store at -20°C.Reduce RNA contamination
Resuspend the bacterial cellsFully suspend the bacterial cells to avoid residual clumpsEnsure uniform lysis
Alkaline lysisAfter adding Solution II, immediately gently invert to mix; do not leave at room temperature for more than 5 minReduce genomic DNA contamination
Neutralize and clarifyGently invert 8–10 times, stand for 2–5 min, and then centrifuge as requiredFully precipitate proteins and impurities
Column washingRepeat washing with Wash Solution once; medium scale also includes W1 washIncrease plasmid purity
Column dryingAfter centrifuging the empty column, open the cap and leave at room temperature for a few minutes to dry completelyAvoid residual ethanol affecting downstream experiments
ElutionSmall scale 50–100 μL, medium scale 100 μL; leave at 37°C for 2 min before centrifugingObtain stable recovery rate
Results detectionTake 1–3 μL for electrophoresis testing; use the product immediately or store at -20°CConfirm the product and store it properly

Applications

From routine cloning screening to multiple batches of downstream experiments, flexibly covering conventional plasmid preparation needs
Positive clone screening

Positive clone screening

Small-scale preparation is suitable for initial screening of recombinants, restriction enzyme analysis, and high-throughput routine screening.

Sequencing and PCR Templates

Sequencing and PCR Templates

High-purity plasmid DNA can be directly used for pre-sequencing preparation and PCR template preparation.

Restriction Ligation and Transformation

Restriction, Ligation, and Transformation

Medium-scale preparation supports multi-tube restriction, large-scale transformation, and multiple batches of molecular cloning experiments.

Cell Transfection and In Vitro Transcription

Cell Transfection and In Vitro Transcription

Relatively large amounts of high-purity plasmid can be used for cell transfection, in vitro transcription, and plasmid preservation.

Frequently Asked Questions

Common issues regarding yield, RNA contamination, lysis, low-copy plasmids, and sequencing quality
What should I do if the plasmid DNA yield is low?
Check whether the bacterial culture is fresh and whether the antibiotic is effective; increase the amount of culture for low-copy plasmids; make sure RNase A has been added to Solution I; the elution volume should not be less than 50 μL (small scale) or 100 μL (medium scale); preheating the Elution Buffer to 65°C can further improve the yield.
What should I do if there is RNA contamination in the product?
Confirm that all RNase A has been added to Solution I; the activity of RNase A solution stored long-term (>3 months) decreases, and fresh RNase A needs to be added; reduce the starting bacterial amount or proportionally increase the volume of lysis buffer.
What should I do if the bacterial solution is not clear after adding Solution II?
Excessive bacterial mass can lead to incomplete lysis, so reduce the amount of bacterial solution or process in two tubes; if Solution I is not fully resuspended, use a vortex to fully suspend it; also check for contamination by exogenous microorganisms.
How to extract low-copy plasmids?
For small kits, increase the bacterial solution to 4–6 mL and centrifuge in batches to combine; for medium kits, increase to 10 mL; proportionally increase the volumes of Solution I/II/III, and pass the supernatant multiple times through the same adsorption column.
What should I do if the sequencing results of purified plasmids are not good?
When the template DNA quantification is inaccurate, it is recommended to use agarose gel electrophoresis for quantification; use fresh bacterial cultures and strictly follow the steps to improve purity; if the insert fragment contains GC-rich, repetitive sequences, or palindromic structures, the sequencing scheme needs to be optimized.

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