Unidade de Genômica Computacional Darcy Fontoura de Almeida (UGCDFA)

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The GS FLX Titanium Sequencing Kits XLR70 and XL+ plus PicoTiterPlate Kit support various bead loading region configurations. The size and number of PicoTiterPlate (PTP) regions to use depend on the sequencing throughput requirements of the experiment, in total bases or total reads. To determine the proper gasket size, proceed as follows:

Use Table 1 to identify the size and number of bead loading regions best suited to yield the sequencing throughput for your DNA library sample. The throughput presented in Table 1 is typical for good 200 cycle (XLR70 kit) or 400 cycle (XL+ kit) sequencing runs performed using good quality shotgun libraries and for amplicon libraries with fragments up to 700 bp. For example, column 4 of Table 1 shows the expected throughput of a full sequencing Run carried out with the longest run script available for this kit (200 cycles, generating an average read length of 350 to 450 nt or 400 cycles, generating reads up to 1 kbp in length), with all regions loaded.

Table 1*: Gasket specification and throughput for shotgun and amplicon libraries.

Region SizeRegions per PTP DeviceBases per Region (Mbp)Bases per Full PTP Device (Mbp)Reads per Region (x103)
XLR70XL+XLR70XL+ShotgunAmplicon
Large2180-280250-400360-560500-800450-650>255
Medium460-110180-250240-440400-640160-250>100
M/S830-5540-70240-440320-56080-120>50
Small1610-2016-25160-320240-40025-40>10
*Adapted from Sequencing Method Manual for the GS FLX+ Instrument (Roche)

Ion PGM e Proton

Compare the data for Ion PGM™ and Ion PI™ Chips, and choose the chip that works best for your sequencing needs:

ChipExpected Sequencing Run TimeExpected Output*Expected Reads
200-base reads400-base reads200-base reads400-base reads
Ion PGM 3142.3h3.7h30-50mb60-100mb400-550K
Ion PGM 3163.0h4.9h300-600mb600mb-1Gb2-3 million
Ion PGM 3184.4h7.3h600mb-1Gb1.2-2Gb4-5.5 million
Ion Proton PI**2-4hUp to 10Gb60-80 million
*Expected output with >99% aligned/measured accuracy. Output is dependent on read length and application
**Up to 200bases fragment reads
Source: Life Technologies

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