Tissue RNA Mini Kit 250 prep R6311-02

Tissue RNA Mini Kit 250 prep R6311-02 kualitas terbaik dan harga murah, produk tersertifikasi kalibrasi Akreditasi KAN. Produk terbaik hanya di BMDStore.COM

Product Description

Buffer LY, RB, RNA Wash Buffer, DEPC-Treated ddH2O, ezBind Columns, Collection Tubes, DNase I (Optional), User Menu

Features :
Tissue,Eukaryotic cells RNA purification miniprep Kit: Purify up to 100μg total RNA from 5×106 Eukaryotic cells or 30mg tissue

Introduction

The EZgeneTM Tissue RNA Kit provides an easy and fast method for isolating total RNA from tissues, cultured cells within 30 min. Only trace genomic DNA exists in the purified RNA, which can be eliminated by DNase I treatment (See detail in the protocol) when it is necessary. This kit purifies up to 100 µg of total RNA from eukaryotic cells or animal tissues. The purified RNA is ready for RT-PCR, Northern blotting, polyA+ RNA purification, nuclease protection, and in vitro translation.

Storage and stability

DNase I (opsional) harus disimpan pada -20 ° C. Semua komponen lainnya dapat disimpan pada suhu kamar. Semua komponen kit dijamin selama 12 bulan sejak tanggal pembelian.

Kit contents

Catalog#

R6311-00

R6311-01

R6311-02

Preps

4

50

250

Buffer LY

2.4 mL

28 mL

135 mL

Buffer RB

3 mL

30 mL

135 mL

RNA Wash Buffer

2 mL

20 mL

3 x 24 mL

DEPC-Treated ddH2O

500 µL

10 mL

30 mL

ezBind Columns

4

50

250

Collection Tubes

8

100

500

DNase I (Optional)*

25 u

300 u

1500 u

DNase Stop Buffer

200 µL

2.4 mL

12 mL

*Solutions are not supplied. They could be purchased from Biomiga.

Safety information

Buffer LY mengandung garam chaotropic, yang dapat membentuk senyawa reaktif jika digabungkan dengan pemutih. Jangan menambahkan pemutih atau larutan asam langsung ke limbah persiapan, kenakan sarung tangan dan kacamata pelindung saat menangani.

Before starting

Persiapkan semua komponen dan persiapkan semua bahan yang diperlukan dengan memeriksa buklet instruksi ini dan menjadi terbiasa dengan setiap langkah.

Important

Tentukan volume Buffer LY yang akan digunakan dan tambahkan 20 µL b-mercaptoethanol (b-ME) per 1 mL Buffer LY sebelum digunakan. Buffer LY berisi b-ME dapat disimpan pada suhu ruangan hingga 1 bulan.
Kristal dapat terbentuk di Buffer LY, larutkan endapan pada suhu 37 oC sebelum digunakan.
Tambahkan 8 mL (R6311-00) atau 80 mL (R6311-01) atau 96 mL (R6311-02) etanol 100% ke RNA Wash Buffer sebelum digunakan. Etanol akhir adalah 80% (v / v).
Opsional: Tambahkan etanol 100% 800 µL atau 9,6 mL (R6311-01) atau 48 mL (R6311-02) ke DNase Stop Buffer sebelum digunakan. Etanol akhir adalah 80% (v / v).

Materials supplied by users

  •  Tabletop microcentrifuge and 1.5 mL sterile tubes.
  •  Vacuum manifold if use vacuum protocol.
  •  100% ethanol.
  •  b-mercaptoethanol.
  •  Optional: DNase I, DNase Buffer

Note: Perform all steps including centrifugation at room temperature

Trouble Shooting Guide

Problems Possible reasons Suggested Improvements
Low A260/A280 ratios

 

Protein contamination Do a Phenol:Chloroform extraction. Loss of total RNA (up to 40%) should be expected.
Low A260/A280 ratios

 

Guanidine Thiocyanate contamination

 

Add 2.5 volumes of ethanol and 0.1M NaCl (final concentration) to precipitate RNA. Incubate for 30 min at -20°C. Centrifuge at 10,000 g for 15 min at 4°C. Resuspend the RNA pellet in DEPC-treated water.
Low Yield RNA in sample degraded

 

Freeze samples immediately in liquid nitrogen and store at -70°C after collect it.
Low Yield The binding capacity of the membrane in the spin column was exceeded Use of too much tissue sample exceeding the binding capacity of spin column will cause the decreasing of total RNA yield.
Low Yield Ethanol not added to buffer Add ethanol to the RNA Wash Buffer and DNase Stop Solution before purification.
Genomic DNA contamination

 

Too much total RNA sample was used in RT-PCR. Reduce total RNA amount used in RT-PCR to 50-100 ng.
Genomic DNA contamination

 

The sample may contain too much genomic DNA.

 

Reduce the amount of starting tissue in the preparation of the homogenate. Most tissues will not show a genomic DNA contamination problem at 30 mg or less per prep.

 

Reduce cell numbers to 1-2×105 or increase buffer volume and do multiple loadings to column.

MORE INFO :
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